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[Clinical efficacy and T-lymphocyte subset, serum interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), interleukin-2(IL-2) levels on treatment of chronic aplastic anemia patients by shenfu injection combined with stanozol and cyclosporin A].

OBJECTIVE: To observe the effect of Shenfu injection (SFI) and influence on T-lymphocyte subset, serum level of interferon-gamma(IFN-gamma), tumor necrosis factor-alpha(TNF-alpha), interleukin-2(IL-2) in patients with chronic aplastic anemia (CAA) based on treating with stanozol and cyclosporin A. METHOD: 60 patients with CAA were randomly divided into two groups, 30 patients in the SFI group were treated with SFI (100 mL which contains Ginsenoside 0.8 mg x mL(-1) and aconitine 1.8 microg x mL(-1) by adding it in 500 mL of 5% glucose every day) plus stanozol and cyclosporin A and 30 patients in the control group treated with slanozol and cyclosporin A alone for 2 months. The clinical efficacy was observed. The change of T-lymphocyte subset analyzed by flow cytometry and the levels of serum IFN-gamma, TNF-alpha, IL-2 measured with ELISA method were also observed before and after treatment. RESULT: After treatment, the total effective rate of the SFI group was higher than that in the control group, but it did not showing significant difference. The CD4/CD8 levels were significantly increased (1.76+/-0.49, P< 0.01) and CD8 levels were significantly lowered (22.57+/-6.30, P < 0.01) in the SFI group after treatment. Serum levels of lFN-gamma, TNF-alpha and IL-2 were lower in both groups, and the level of TNF-alpha and IL-2 in the SFI group (0.710+/-0.213) ng x L(-1) and (0.639+/-0.247) ng x L(-1) was significantly lowered than that in the control group (P < 0.05, P < 0.01). CONCLUSION: SFI might believe the hemopoietic inhibition so as to promote the recovery of hemopoietic function through improving the T-lymphocyte subset and reducing the release of hemopoietic negative regulatory factors such as IFN-gamma, TNF-alpha and IL-2.

Aconitine↗

Mitoxantrone as a single agent in pretreated metastatic breast cancer: effects on T lymphocyte subsets and their relation to clinical response.

Mitoxantrone (DHAD), an anthracenedione with antineoplastic properties similar to doxorubicin, was tested for therapeutic efficacy and for immunomodulating action on lymphocyte subsets in 16 metastatic breast cancer patients, 12 of whom had been previously treated with chemotherapy. DHAD was given intravenously at a dose of 14 mg/m2 every 21 days. To evaluate total T lymphocytes (CD3), T helper (CD4), and T suppressor/cytotoxic cells (CD8) and the CD4/CD8 ratio, venous blood samples were drawn before and after the first DHAD cycle. Moreover, in 8/16 patients, B lymphocytes (CD20), T suppressor cells (CD8+/CD57+), T cytotoxic cells (CD8+/CD57-), NK (CD16) and IL-2 receptor-expressing cells (CD25) were also measured at the same time. An objective tumor response was achieved in 5/16 (31%) patients and the response rate was significantly higher in patients pretreated with hormone therapy alone than in those pretreated with chemotherapy. No relation was found between clinical response and changes in the CD4/CD8 ratio. Neither the mean number nor the percentage of CD3, CDA and CD8 cells observed after DHAD were significantly different with respect to those seen before. In contrast, the mean number of T suppressor cells, B lymphocytes and CD25-positive cells was significantly lower after than before DHAD administration, whereas no difference was seen in NK cells. These results confirm in humans the immunomodulating properties of DHAD previously described in experimental conditions. However, the DHAD-induced changes in lymphocyte subsets do not seem to be related to the clinical response in breast cancer.

Adult↗

Lymphocyte subsets and sister-chromatid exchanges in the students exposed to formaldehyde vapor.

The present report evaluates the effects of formaldehyde (FA) exposure on peripheral lymphocytes by using both genetic and immunological parameters. Twenty-three non-smoking students in the study had inhalation exposure to 0.508 +/- 0.299 mg/m3 of FA for a period of 8 weeks (3h x 3 times each week) during anatomy classes. As for composition of lymphocyte subsets after FA exposure, significant increase was found in the percentage of CD19 (B cells), while significant decrease was observed in CD3 (total T cells), CD4 (T helper-inducer cells), and CD8 (T cytotoxic-suppressor cells) with a P < 0.01. Increase in the ratio of T-helper-inducer cells to T-cytotoxic-suppressor cells (T4/T8) was also observed with statistical significance after exposure (P < 0.001). In the meanwhile, no significant difference (P > 0.05) was reported between lymphocyte proliferation rate and sister-chromatid exchange (SCE) at the exposure level and duration. It is suggested that the lymphocyte subsets may be most susceptible to the effects of FA, though a single immunological endpoint is rarely related with pathophysiological interpretation.

Adult↗

Lymphocyte subset analysis and glycosylphosphatidylinositol phenotype in patients with paroxysmal nocturnal hemoglobinuria.

Using multicolor flow-cytometry we have examined 19 patients with paroxysmal nocturnal hemoglobinuria (PNH) (18 with active disease and 1 spontaneous remitter) to determine absolute numbers of lymphocyte subsets and the proportion of glycosylphosphatidylinositol (GPI)-deficient clones amongst these subpopulations. Lymphocyte subsets were abnormal in all patients; the most frequent findings were low absolute numbers of natural killer (NK) cells (median, 0.08 x 10(9)/L; normal range, 0.2 to 0.4 x 10(9)/L) and low absolute numbers of B cells (median, 0.05 x 10(9)/L; normal range, 0.06 to 0.65 x 10(9)/L). GPI-deficient B, T, and NK cells were identified in 88%, 84%, and 89% of patients, respectively. The proportion of GPI-deficient cells within individual lymphoid lineages was highly variable, though in most patients the percentage of GPI-deficient NK cells was considerably higher than B or T cells. These observations can be explained when mechanisms of normal lymphopoiesis are considered. Despite these quantitative and qualitative abnormalities, no patients suffered an excessive number or severity of infections. The detection of PNH clones amongst all lymphocyte lineages may provide important information regarding the natural history of the disease and additional insights into kinetics of adult lymphopoiesis.

Adult↗

Blood transfusions and immunophenotypic alterations of lymphocyte subsets in sickle cell anemia. The Transfusion Safety Study Group.

Transfusions purportedly induce dysfunction of cell-mediated immunity in sickle cell anemia (SCA). We studied hematologic and lymphocytic indices in 173 human immunodeficiency virus (HIV)-negative subjects with SCA and 131 black controls. Children aged 1 to 7 years with SCA had leukocyte counts and percentages of granulocytes, monocytes, natural killer cells, and T-cell markers (CD2+CD11b+, CD4+CD26+, CD4+CD29+) that were significantly higher than those for control children. Percent total lymphocytes was decreased for this age group, but the total number of lymphocytes and T and B cell counts were similar to controls. Platelets were not increased. Adolescents (aged 8 to 17 years) and adults (aged > or = 18 years) with SCA had increased total leukocytes and monocytes and lymphocytes counts that remained level instead of decreasing, as did comparably aged controls. Lymphocyte subsets typically increased in count, but their percentage remained similar to children. The exception was CD56+ cell counts, which were increased in adolescents and adults. No lymphocytic subset change suggested impaired cellular immunity, and none could be related to transfusion. Prophylactically transfused patients had higher granulocyte counts, but these may arise from the complications of SCA itself.

Adolescent↗

Circulating intercellular adhesion molecule-1 and lymphocyte subsets involved in immune response of breast cancer.

To understand the immune responses of patients with breast cancer, circulating intercellular adhesion molecules-1 (ICAM-1) and lymphocyte subsets were measured in patients with breast cancer and in healthy persons. There were 10 patients of with metastasis, 11 patients breast cancer without metastasis and 9 age-and-sex matched healthy persons. Their serum samples were counted for circulating ICAM-1 levels, and blood cells were calculated for lymphocyte subpopulation including CD3, CD4, CD8 and CD19 by using flow cytometry. Higher levels of circulating ICAM-1 were noted in patients with breast cancer and metastasis than those without metastasis (p < 0.01). Lower percentage of CD8 lymphocyte subset were found ih patients with breast cancer and metastasis than those without metastasis (p < 0.05). Shedding of ICAM-1 antigens from the tumor may block the attachment of cytotoxic T cells, and decreased numbers of cytotoxic T cells may be unable to inhibit the spread of cancer cells. Immune responses of patients with breast cancer may be different between metastatic status and limited status. Further investigations should correlate the results of tissue stains of breast cancer with ICAM-1 and lymphocyte markers with those from peripheral blood.

Adult↗

Lymphocyte subsets in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is an autoimmune disease characterised by increased B cell activity and depressed T cell function. However, the contribution of the immunoregulatory system to its pathogenesis is still unclear. The recent development in the production of monoclonal antibodies and the availability of bench-top flow cytometers have allowed rapid quantitation of peripheral blood lymphocyte subsets. We analysed the distribution of the lymphocyte subsets in 24 patients with active SLE and 18 with inactive SLE. The distribution of immunoregulatory cells in 72 normal volunteers was used as control. Statistical analysis showed that there were significant differences between both the SLE groups and the normal controls, for total lymphocytes, T cells, B cells, T helper cells, T suppressor cells, T helper/suppressor ratio and natural killer cells. There was a significant difference for T helper cells between active and inactive SLE. T helper cells levels were found to be low in inactive SLE and lower in active SLE. It appears that treatment-induced remissions did not restore the levels of immunoregulatory cells to normal. Thus, T helper cell levels reflect disease activity and longitudinal assays of T helper cells may serve as an indicator of disease reactivation.

Adult↗

Implications of circulating autoantibodies and peripheral blood lymphocyte subsets for the genesis of premature ovarian failure.

Several kinds of circulating autoantibodies and peripheral blood lymphocyte subsets were studied in 20 patients with secondary amenorrhea manifesting hormonal and clinical features of premature ovarian failure (POF). More than one kind of autoantibody was detected in 14 patients (70%). Seven patients (35%) had anti-thyroglobulin antibody, 6 (30%) had anti-parietal cell antibody, 8 (40%) had anti-nuclear antibody, and one patient with chronic thyroiditis had anti-TSH receptor antibody. Anti-adrenal cortex antibody and RA test were negative in all patients. Two patients had clinically evident autoimmune disease; one had myasthenia gravis and the other had chronic thyroiditis. Examination of peripheral blood lymphocyte subsets of 19 patients by flow cytometry revealed an increase in the percentage of OKT3+ cells and OKT4+ cells and a decrease in OKT8+ cells in POF patients compared with age-matched controls, but these differences were not significant. An increase in the OKT4/OKT8 ratio was, however, significant. It has been suggested that an autoimmune mechanism may participate in the genesis of POF, at least in patients with autoimmune diseases; however, the findings in this study support the hypothesis that some pure POF may also be caused by an autoimmune process resulting from a subclinical imbalance in the immunoregulatory system before manifestation of the autoimmune disease.

Adult↗

The association between CD2+ peripheral blood lymphocyte subsets and the relapse of bladder cancer in prophylactically BCG-treated patients.

We investigated the potential existence of differences in the distribution of T-lymphocyte subsets and in the proliferative response of these CD2+ cells to polyclonal mitogens in patients with transitional cell bladder carcinoma (SBTCC) treated with prophylactic intracavitary instillations of bacillus Calmette-Guerin (BCG) according to their clinical response to this treatment. Before BCG treatment, different subset distribution (CD8+ and CD3+ CD56+), activation antigen expression (CD3+ HLA- DR+) and proliferative response to mitogenic signals were found in CD2+ cells from SBTCC patients prophylactically treated with BCG who remained free of disease or those who had recurrence of tumour. Otherwise, the prophylactic intracavitary BCG instillations in SBTCC patients are associated with a transitory variation of T-lymphocyte subset distribution (CD4 and CD8) and activation antigens expression (CD25).

Adjuvants, Immunologic↗

Lymphocyte subsets in Bell's palsy: immune pathogenesis and outcome prediction.

The aim of this prospective study is to define the prognostic significance of lymphocyte subset analysis in children with Bell's palsy. Lymphocyte subgroup analysis in peripheral blood was performed in 17 children with Bell's palsy by using flow cytometry. Before a standard protocol of corticosteroid treatment, patients were categorized into two groups for facial nerve impairment on the basis of the clinical findings: Group 1 (mild to moderate impairment), 7 patients; and Group 2 (severe impairment), 10 patients. Outcome of the patients was evaluated at the end of 3 months follow-up and categorized as satisfactory recovery (n = 12) or unsatisfactory recovery (n = 5). Decreased percentages of B cells (CD19) and T helper/inducer (CD4) subsets were measured in patients with Bell's palsy compared with age-matched healthy control patients. Patients with severe impairment had significantly lower percentages of CD4 and CD19 subsets, whereas patients with mild to moderate impairment had only decreased percentage of CD19 subsets. There was no statistically significant difference in the percentage of lymphocyte subsets between the patients with satisfactory and unsatisfactory recovery. These results provide additional support for cell-mediated immunopathogenesis in patients with Bell's palsy, without any prognostic significance for the outcome.

Adolescent↗

Melatonin restores and enhances the human type B tonsillar lymphocyte subset in recurrent acute tonsillitis.

Tonsils have a privileged situation in the immune system in that they are in touch with the environment. Melatonin is a hormone that is influenced by the circadian environmental variations of dark-light and is a modulator of the immune system. We have studied a group of thirty five children with recurrent acute tonsillitis that were submitted for tonsillectomy. Tonsillar lymphocyte subsets were determined before and after culture through flow cytometry in a tonsillar mononuclear suspension. After the culture, the lymphocyte subsets of type B suffered a decrease that was restored in the presence of melatonin or phytohemaglutinin, and even increased above the values of the control when the culture was accomplished in the presence of both substances. This process was specific for B cells, no occurrence for T lymphocytes or natural killer cells. Melatonin is found in the crossroads of the interaction of the microorganisms, pollens or inert substances with the tonsillar lymphocytes in the production of the immune defences. Further study is required on tonsillar pathology to explain its physiopathology and its possible therapeutic role.

Acute Disease↗

Lymphocyte subset distribution and cytokine secretion in third trimester decidua in normal pregnancy and preeclampsia.

BACKGROUND: Excessive Th1 activity in peripheral blood plays a probable role in the pathogenesis of preeclampsia. The aim of the study was to investigate whether disturbed local immune reactions are also present in decidua. METHODS: Flow cytometric analysis of CD3, CD19, CD56/CD16, CD4, CD8, CD4/CD29, CD4/CD45RA, CD4/CD45RO, CD8/CD28, CD3/CD69 lymphocyte subsets isolated from third trimester decidua of pregnants with preeclampsia (n=21) and pregnant controls (n=11) subjected to elective caesarean sections. Spontaneous and phytohemaglutynine stimulated "in vitro" secretion of IL-2, IL-4, IL-6, IL-10, IL-12, IFN-gamma and TGF-beta by decidual lymphocytes was studied by ELISA. For the statistical significance of differences between the groups the U Mann-Whitney test was performed (confidence interval P<0.05). RESULTS: Preeclamptic patients were characterized with an increased percentage of the CD3-/CD56+CD16+, CD8+/CD28+ and decreased percentage of CD3+, CD19+, CD4+/CD45RA+ lymphocytes. The profile of secreted cytokines shifts in favor of Th1 activity (extremely high IFN-gamma and low IL-6 and IL-10 secretion). Decidual IL-12 secretion in preeclamptic patients is decreased compared to controls. CONCLUSION: Changes in NK and T lymphocyte subsets followed with Th1 cytokine IFN-gamma over-activity, could affect local immunoregulatory mechanisms in third trimester decidua of preeclamptic patients.

Antigens, CD↗

Identification of lymphocyte subsets in peripheral blood and thyroid gland in Graves' disease by alpha-naphthyl-acetate-esterase reaction.

T lymphocyte subsets were prospectively examined in the peripheral blood and thyroid aspirates of 10 patients with hyperthyroid Graves' disease before and after treatment with methimazole and attainment of euthyroidism. T lymphocyte subsets were identified with monoclonal antibodies and pattern of alpha-naphthyl-acetate esterase (ANAE) staining pattern in the case of peripheral blood and ANAE staining pattern with thyroid aspirate smears. Before treatment, OKT8+ lymphocytes were significantly decreased (18.4% +/- 4.8) (S. D.) in the patients compared to control (28.8 +/- 6.7%, p less than 0.05), the OKT4/OKT8 ratio was increased (2.92 vs 2.11). Percent OKT8+ lymphocytes were not different from the controls when the ten patients had been rendered euthyroid. ANAE mononuclear cells with a diffuse pattern (presumed suppressor cells) were 4.2% +/- 1.8 before treatment and 8.3 +/- 2.4 (p less than 0.05) after treatment and 11.5% +/- 2.2 in controls. ANAE mononuclear cells with diffuse pattern represented 4.2% +/- 1.8 of the mononuclear cells infiltrating the thyroid gland of untreated patients and rose to 8.3% +/- 2.4 after the patients had become euthyroid. ANAE negative cells (B cells and some T cells) were increased in the thyroid of untreated patients. It is concluded that mononuclear cells with presumed suppressor T cell phenotype are decreased in the blood and thyroid glands of patients with active Graves' disease and that this defect is corrected when euthyroidism has been established.

Adult↗

[Dynamics of T lymphocyte subsets in HAART treated AIDS patients with successful suppression of HIV replication and different CD4 + T cell restoration].

OBJECTIVE: To study the dynamic changes of T lymphocyte subsets of AIDS patients during more than 24 months of highly active antiretrovirus therapy (HAART) with successful suppression of HIV replication and different CD4 + T cell restoration. METHODS: Totally 45 AIDS patients who had received HAART for more than 24 months were included. During HAART (including DO, M3, M6, M12, M18, and M24), the number of plasma HIV-1 RNA was measured quantitatively using the bDNA assay, and T lymphocyte subsets including CD3 + CD4 + cells, CD3 + CD8 + cells, naive CD4 + cells (CD4 + CD45RA + CD62L +), CD4 + CD28 + cells , and CD8 + CD38 + cells were detected with flow cytometer. RESULTS: Among 45 patients, 24 patients (53.3%) whose plasma viral load decreased to less than 500 copies/ml at M6 and maintained to M24 were classified into three groups according to the CD4 + T cell count increments on M24 (compared with DO): group A (< 100/mm3), group B (100-200/mm3), and group C (> 200/mm3). After the initiation of HAART, T lymphocyte response, including CD4 + T cell counts, naive CD4 + cell counts, percentages of CD4 + CD28 + cells in these patients were improved gradually, while CD8 + CD38 + percentage decreased. The improvement of T lymphocyte response in group C was most remarkable even with highest plasma viral load and lowest CD4 T cell count on DO. Compared with group A and B, group C had significantly better improvement not only in the quantities of CD4 + T cell, but also in the CD28 + expression and naive CD4 + T cell populations. CONCLUSIONS: T lymphocyte response of AIDS patients can be effectively reconstituted by HAART. Different dynamics of CD4 + CD28 + and naive CD4 + populations may considerably contribute to the quantity and cellular function restoration of CD4 + T lymphocyte.

Acquired Immunodeficiency Syndrome↗

[Determination of lymphocyte subsets in pulmonary tuberculosis by Mc-Ab- rosette reagent method].

The lymphocyte subsets in peripheral blood were determined in 73 cases of active pulmonary tuberculosis and 40 healthy subjects using rosette reagent of monoclonal antibody (McAb) to anti-T lymphocyte. The results showed that the percentages of T3, T4, T8, and T4/T8 ratio were 54.3 +/- 6.7%, 34.3 +/- 7.9%, 35.5 +/- 10.0%, 1.0 +/- 0.4 of patients group 66.3 +/- 7.9%, 45.3 +/- 6.3%, 29.9 +/- 5.8% and 1.6 +/- 0.3 of healthy control group respectively. There was statistically significant difference between these two groups (P less than 0.01).

Adult↗

T-lymphocyte subsets in patients with hookworm infection in Zaria, Nigeria.

Peripheral lymphocyte subsets CD3, CD4, CD8 were studied using monoclonal antibodies to determine the mechanism of immunosuppression observed in an earlier study with total T-cells using the sheep erythrocyte rosetting technique. The study was carried out in 37 Nigerian patients with hookworm infection (Necator americanus and Ancylostoma duodenale and 17 healthy Nigerians as controls. The leucocyte migration inhibition (LMI) test was also carried out to assess the functional integrity of the lymphocytes, while the hookworm status of the patients was assessed by the Stoll technique for measuring egg counts. The results of the T-cell studied showed that CD3 and CD4 cell percentages were significantly depressed in hookworm patients compared to controls (P<0.05). The difference between mean CD8 percentages and absolutes in hookworms patients and controls were not significant. Absolute CD3 and CD4 numbers were not significantly lowered compared to controls in the study and CD4/CD8 ratios were insignificantly reduced. Patients with polyparasitism including hookworm infection showed significant reduction in CD3 and CD4 cells when compared with controls and with patients with hookworm infection alone. The leucocyte migration inhibition response to purified protein derivative of M. tuberculosis (PPD) was significantly decreased in hookworm patients compared to controls, confirming that functionally cell mediated immunity is depressed in hookworm infection.

Adolescent↗

Dual-color flow cytometric analysis of phenotype, activation marker expression, and proliferation of mitogen-stimulated bovine lymphocyte subsets.

Bovine peripheral blood mononuclear cells were cultured in vitro for 3 days with the mitogens concanavalin A (Con A), pokeweed mitogen (PWM), phytohemagglutinin (PHA), and anti-CD3 monoclonal antibody. Activation of T-lymphocyte subsets (CD4+, CD8+, and gammadelta T-cell receptor+) and of B-cells was measured by two-color flow cytometric analysis of subset expression of IL-2 receptor alpha (CD25) and MHC class II. Proliferation of lymphocyte subsets was directly measured by two-color flow cytometric analysis of fluorescence intensity of PKH2, a fluorescent dye that stably incorporates into cell membranes. CD4+ and CD8+ T-cell subsets were stimulated by all the mitogens to increase expression of IL2r and MHC II and to proliferate. Deltagamma+ T-cells responded to all four mitogens with increased IL2r and MHC II expression. Con A and PHA caused measurable proliferation of deltagamma+ T-cells, but PWM and anti-CD3 did not. B-cells generally responded to the mitogens with increased IL2r and MHC II expression. B-cells proliferated when incubated with Con A, but did not measurably proliferate in response to PWM, PHA, or anti-CD3.

Animals↗

A comparison of lymphocyte subset distribution in rat lacrimal glands with cells from tissues of mucosal and non-mucosal origin.

The subset distribution of lymphocyte populations isolated from rat lacrimal glands (LG) was compared with those from tissues of both mucosal and non-mucosal origin. In spleen (SPL), mesenteric (MLN) and cervical lymph node (CLN) populations the percentages of cells bearing W3/13 (pan T) and OX19 (pan T) were greater than the percentages obtained for cells bearing the OX7 (Thy-1) marker. In contrast, for lacrimal (LG), salivary (SG) and mammary gland (MG) populations, cells bearing OX7 predominated over those bearing the W3/13 and OX19 markers, with the exception of day 1 post-partum MG tissue which displayed equal numbers of OX7 and OX19 cells. Except for MG, in which OX8 (T non-helper) predominated over W3/25 (T helper) populations, the proportions of these two subsets in the other tissues were generally similar. Analysis of SPL and LG cells for coexpression of OX7 with OX19 or L chain indicated that significant percentages of OX7 bearing cells also expressed T or B cell markers. However, the higher values noted for the OX7 population in LG were not attributable to increased numbers of cells coexpressing pan T or B cell markers. These findings show that lymphocyte subset distribution in LG and other glandular mucosal tissues is distinct from that of non-mucosal tissues, in that mucosal tissues contain a predominance of cells bearing the Thy-1 (OX7) phenotype.

Animals↗