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PubMed2003

Flow-cytometric analysis of T-lymphocyte subsets after different treatment methods in smokers and non-smokers with chronic periodontitis.

AIM: To determine any change in T-lymphocyte subsets after applying different treatment methods in smokers and non-smokers with chronic periodontitis. PARTICIPANTS: 50 adults with chronic periodontitis. METHOD: The subjects were divided into smokers and non-smokers. Biopsy samples were taken from the gingival pocket wall tissues at sites with chronic periodontitis before treatment, after initial treatment, after curettage and after flap operation and tested for CD4+, CD8+ lymphocyte and CD4/ CD8 ratio values. Gingival pocket depth, gingival index (GI-Löe-Silness) and plaque index (PI-Silness-Löe) scores were also recorded. Analysis aimed at determining the relation between the clinical measurements and the laboratory results. RESULTS: Flow cytometry findings in both groups showed that CD4+ and CD8+ lymphocyte values before treatment were under the normal value while the CD4+/CD8+ ratio was within normal distribution interval. The lymphocyte values observed in the smokers were found to be lower than those in the non-smokers. After treatment the difference between the lymphocyte values in smokers and non-smokers was found to be statistically significant. However, the difference between the CD4/CD8 rate obtained in smokers and non-smokers was not found to be statistically significant. CONCLUSIONS: The lymphocyte values observed in smokers were found to be lower than those in non-smokers after applying different treatment methods and the local immune response was poor in the smokers.

Adult↗
PubMed2000

[An experimental study of immune mediating action of T-lymphocyte subsets on recurrent herpetic stromal keratitis].

OBJECTIVE: To investigate the role of T-lymphocyte subsets in the recurrent herpetic stromal keratitis (SK). METHODS: In a recurrent SK NIH mouse model, rat monoclonal antibodies were used to selectively deplete in vivo the CD(+)(4) and CD(+)(8) T-lymphocytes in mice, and the effects of the subsets on recurrent herpetic SK were evaluated. RESULTS: As measured by flow cytometry, in the mice treated with GK1.5, > 90% of CD(+)(4) T cells were depleted; and in the mice treated with H-35, > 95% of CD(+)(8) T cells were depleted. Depletion of CD(+)(4) T cells could halt the recurrence of SK, reduce the severity of recurrent SK and impair the delayed type hypersensitivity. In contrast, depletion of CD(+)(8) T cells had no effect on the outcome of the disease. Histopathologic examination of the recurrent SK demonstrated that the stromal inflammation in CD(+)(8) T-cell depleted and control mice was characterized by extensive lymphocyte and polymorphonuclear leukocyte infiltration, but only occasional lymphocytes were observed in CD(+)(4) T-cell depleted mice. CONCLUSION: The results provide evidence that recurrent herpetic SK represents an immunopathologic process mediated by CD(+)(4) T cells.

Animals↗
PubMed1997

Alterations of lymphocyte subsets in patients with recurrent fetal wastage positive for antiphospholipid antibodies treated with Chinese herbal medicine.

The alterations of peripheral blood lymphocyte subsets were sequentially analyzed in patients with recurrent fetal wastage who were treated with the Chinese herbal medicine, Sairei-to (Chan ling-Tan) for positive antiphospholipid antibodies to analyze the underlying mechanisms of the therapy. The titer of antiphospholipid antibodies was significantly decreased by administration of Sairei-to at one and two months after commencement of treatment and in the newly pregnant state compared with that before administration. The percentage of CD19-positive cells significantly decreased at two months after commencement of Sairei-to treatment and at the newly pregnant state compared with that before administration in successful pregnancies. The percentage of CD4-positive cells significantly increased two months after commencement of Sairei-to treatment compared with that before administration in both successful pregnancies and total cases. The CD4/CD8 ratio was increased significantly after two months administration of Sairei-to in both successful pregnancies and total cases. Thus, it is suggested that the administration of Sairei-to might induce the predominance of CD4-positive cells in parallel with the suppression of antiphospholipid antibodies. Moreover, the suppression of CD19-positive cells (B cells) was induced by administration of Sairei-to which might be involved in successful continuation of pregnancy in the patients.

Abortion, Habitual↗
PubMed1992

Modulation of lymphocyte subsets due to psychological stress in patients with rheumatoid arthritis.

The influence of a 1-hour neuropsychological stress test on the distribution of lymphocyte subpopulations and on plasma catecholamine levels was investigated in 18 patients with rheumatoid arthritis (RA) and 14 sex- and age-matched controls. Despite significant increases in lymphocyte counts in both groups, lymphocyte subsets did not change accordingly. A wide scattering of catecholamine levels in plasma before and after stress was observed. Plasma levels of lymphokines such as interleukin (IL)-1 beta and IL-6 could not be detected in RA patients. Enzyme immunoassay of markers of lymphocyte activation such as HLA-DR and cell-bound IL-2 receptor showed only a significant elevation of HLA-DR marked cells in RA patients at baseline. Significantly higher amounts of the soluble IL-2 receptor were detected in patients with RA before the stress test, but stress testing did not alter this parameter. In conclusion, lymphocyte activation in RA and a defect in the expression of IL-2 receptor on the cell surface of lymphocytes were confirmed in the present study.

Arthritis, Rheumatoid↗
PubMed2006

Lymphocyte subset distribution in apparently normal and single intradermal test-positive water buffaloes analyzed by flow cytometry.

Monoclonal antibodies (mAbs) against bovine lymphocyte cell surface antigens namely, MHC Class I, MHC class II (DP, DQ and DR), CD3, CD4, CD8, gamma delta TCR, WC1N1 and WC1N2, were tested for their reactivity on apparently normal buffalo mononuclear cells prepared from spleen, lymph nodes and peripheral blood. All the mAbs cross-reacted with the buffalo mononuclear cells. The mean (+/-SD) CD4:CD8 cell ratio in the peripheral blood of apparently normal buffaloes was 1.08+/-0.049 while in the spleen and lymph nodes it was 0.90+/-0.080 and 1.81+/-0.430, respectively. The lymphocyte subsets in the buffaloes positive for tuberculosis by the single intra dermal (SID) test was found to be altered; the CD4 cells were reduced while the CD8 and gamma delta cells were increased. The mean CD4:CD8 ratio in the SID positive buffaloes was 0.36+/-0.010.

Animals↗
PubMed1986

Quantitative study of the migration of lymphocyte subsets through 3-D collagen gels.

Migratory properties of normal peripheral blood lymphocyte subsets were studied with a 3-D collagen gel model. Both the T and B subsets entered the gel at the same rate, but once inside the T subsets migrated significantly faster than the B subsets. In the T subsets, both the T-helper and T-suppressor cells entered the gel at the same rate, but the former migrated significantly faster once inside the gel. The biological significance of their migrations is discussed.

B-Lymphocytes↗
PubMed1986

[Lymphocyte population and T lymphocyte subsets in brucellosis analyzed by monoclonal antibodies].

In this study, T-lymphocyte subsets and HLA-DR antigen (+) cells in the peripheral blood of 19 patients with acute brucellosis were determined and the results were compared to 19 healthy control subjects. Our findings suggest that exposure of the immune system to brucella microorganism and related antigens results in an increase in the circulating "activated" cell numbers (HLA-DR+) without significant alterations in the subpopulations of peripheral blood lymphocytes.

Acute Disease↗
PubMed2003

Effect of cadmium on lymphocyte subsets distribution in thymus and spleen.

This work was designed to analyze the possible dose dependent effects of cadmium on the distribution of lymphocyte subsets within the thymus and spleen. Cadmium accumulation was also evaluated in these tissues. For this purpose, adult male rats were exposed for one month to 0, 5, 10, 25, 50 or 100 ppm of cadmium chloride (CdCl2) in the drinking water. In both spleen and thymus, the B lymphocytes increased with the doses of 5 and 10 ppm of CdCl2, and decreased with the doses of 25-100 ppm. In spleen, the doses of 25 and 50 ppm decreased CD4+ cells and the doses of 5 and 10 ppm increased CD8+ cells, while the percentage of thymus T, CD4+, CD8+ and CD4(+)-CD8+ cells was not modified by cadmium treatment at any dose used in this study. After cadmium exposure, the metal was accumulated in the spleen only from the dose of 50 ppm on, and in the thymus, from the dose of 10 ppm on. In conclusion, although the accumulation of the metal is higher in thymus than in spleen, the metal affected CD4+ and CD8+ lymphocytes at the spleen but not at the thymus.

Animals↗
PubMed2000

Characterization of lymphocyte subsets in rhesus macaques during the first year of life.

Establishing reliable phenotypic data sets from the analysis of peripheral blood lymphocytes of normal animals is required to assess disease states. The rhesus macaque animal model is well established with respect to adult animals, but limited data are available that characterizes lymphocyte subsets in normal neonates. To address this, we used four-color flow cytometric analysis to follow phenotypic changes in 29 normal rhesus animals through their first ten months of life. From birth to 44 wk of age, the white cell count and absolute lymphocyte count were both elevated compared to adults. CD4+ cells constituted over 80% of all T cells at birth, a percentage that declined gradually over the first 12 wk of life, coincidental with increases in the percentages of CD8+ T cells, CD3-8+ natural killer cells and CD20+ B cells. This difference in relative frequency of CD4 and CD8 results in a significant skewing of CD4:CD8 ratio from 0.7:1 in adults to 3.5:1 in neonates. In addition, the predominant population of T lymphocytes consisted of CD45RA+CD62L+ naive cells. This subset continues to be the predominant phenotype for at least the first year of age. After birth the expression of activation markers (CD25) increased particularly on CD4+ T cells, although these levels generally reached a frequency similar to that observed in adults between 12 and 20 weeks after birth. These results are similar to those seen in humans and further confirm the reliability of the rhesus macaque animal model to study human diseases.

Age Factors↗
PubMed2000

Analysis of some peripheral blood lymphocyte subsets in relation to Borrelia burgdorferi antibodies in patients with Lyme disease.

The aim of our study was to evaluate the changes in T (CD3), B(CD19) lymphocytes, CD4 and CD8 subsets, activated CD3+ HLA-DR+ lymphocytes, lymphocytes with receptor for IL-2 (CD3+CD25), NK cells as well as the rate of CD4/CD8 in 30 patients with recognized Lyme disease, before and after antibiotic therapy. Patients were divided into the following groups: Group I n = 9--without detected specific antibodies against B. burgdorferi and with clinical recognition of EM(erythema migrans). Group II n = 10--with increased IgM production, with a clinical form of Lyme arthritis and neuroborreliosis. Group III n = 11--with increased IgG anti B. burgdorferi production, with clinical recognition of Lyme arthritis. The results were compared with the results obtained in the control group consisting of 90 healthy people. The measurements were performed in the flow cytometer COULTER EPI XL with Becton Dickinson antibodies. The antibodies against B. burgdorferi were detected by means of ELISA method using Dako, Biomedica and Biocom kits. The statistic analysis was performed with AnStat Program. The changes in lymphocyte subsets were characterised by the decrease in the percentage of CD4, CD8, NK and CD3+HLA-DR+ lymphocytes in peripheral blood before treatment with later tendency to increase. The results show that the lag phase of antibodies production coincides with high activity of lymphocytes (group I). The significant level of antibodies in IgM class induced by the collaboration of T and B cells was found in the group II. In the group III, in which antibodies in IgG class predominated, the changes in lymphocyte subsets were less intensive. The results of our investigations indicate that the immune response in Lyme disease develops mainly with the participation of both cellular and humoral response which is involved in both the defense against and the pathogenesis of the disease.

Adolescent↗
PubMed1997

Lymphocyte subsets in patients with ovarian and breast cancer.

Peripheral blood lymphocytes (PBL) from patients with ovarian or breast cancer or benign lesions of the breast, respectively, have been analysed for expression of phenotypic and activation markers by flow cytometry. The results were compared with those of a control group of healthy women. The relative proportion as well as the absolute counts of B lymphocytes were similar in both groups and in the control group. The absolute number of T cells was decreased in breast cancer patients (p < 0.05). The CD4+/CD8+ ratio was significantly depressed in ovarian cancer patients (p < 0.05), but not in breast cancer patients. In the ovarian cancer group, the percentage of CD3+ T cells expressing HLA-DR (p < 0.05) as well as CD3+ T cells expressing CD16 and CD56 (p < 0.05) was significantly higher. The relative proportion as well as the absolute counts of CD3+ T cells expressing the IL-2 receptor (CD25) were significantly higher (p < 0.001), respectively, in breast cancer patients (p < 0.05). These results suggest that gynaecological cancer is associated with specific alterations in the T cell population.

Adult↗
PubMed1986

Methodological considerations for implementation of lymphocyte subset analysis in a clinical reference laboratory.

As the diagnostic utility of lymphocyte subset analysis has been recognized in the clinical research laboratory, a wide variety of reagents and cell preparation, staining and analysis methods have also been described. Methods that are perfectly suitable for analysis of smaller sample numbers in the biological or clinical research setting are not always appropriate and/or applicable in the setting of a high volume clinical reference laboratory. We describe here some of the specific considerations involved in choosing a method for flow cytometric analysis which minimizes sample preparation and data analysis time while maximizing sample stability, viability, and reproducibility. Monoclonal T- and B-cell reagents from three manufacturers were found to give equivalent results for a reference population of healthy individuals. This was true whether direct or indirect immunofluorescence staining was used and whether cells were prepared by Ficoll-Hypaque fractionation (FH) or by lysis of whole blood. When B cells were enumerated using a polyclonal anti-immunoglobulin reagent, less cytophilic immunoglobulin staining was present after lysis than after FH preparation. However, both preparation methods required additional incubation at 37 degrees C to obtain results concordant with monoclonal B-cell reagents. Standard reagents were chosen on the basis of maximum positive/negative separation and the availability of appropriate negative controls. The effects of collection medium and storage conditions on sample stability and reproducibility of subset analysis were also assessed. Specimens collected in heparin and stored at room temperature in buffered medium gave reproducible results for 3 days after specimen collection, using either FH or lysis as the preparation method. General strategies for instrument optimization, quality control, and biohazard containment are also discussed.

Antibodies, Monoclonal↗
PubMed2004

[Difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia].

BACKGROUND: To clarify the difference and significance of T-lymphocyte subsets in differential diagnosis between severe acute respiratory syndrome SARS) and common atypical pneumonia. METHODS: Totally 100 patients hospitalized in Beijing Ditan Hospital since March to June 2003 with clinical diagnosis of SARS were involved in this study. These patients courses of disease were over 3 weeks. These patients were divided into two groups, SARS group and common atypical pneumonia group (non-SARS group). The counts of CD3+, CD4+ and CD8+ T-lymphocyte of two groups were systematically recorded and analyzed. RESULTS: Sixty-five of the patients were confirmed to have common type of SARS, including 26 males and 39 females, 50 cases received methylprednisolone treatment. Thirty-five cases had common atypical pneumonia (non-SARS), 21 were males while 14 were females, 20 cases received methylprednisolone treatment. All the cases of two groups were cured in the end. The SARS patients T-lymphocyte counts decreased first and then increased. Before 15 days of disease course, mean CD3+, CD4+, CD8+ T-lymphocyte counts of SARS patients were decreased apparently (694+/-568/microl, 441+/-356/microl, 309+/-462/microl). After 15th day of disease course, the counts gradually returned to normal CD3+, CD4+, CD8+ T-lymphocyte counts of non-SARS patients were normal. Compared with patients of the same group who were not treated with glucocorticoids, T-lymphocyte counts of non-SARS patients treated with glucocorticoids had no obvious difference. But glucocorticoids had some effect on SARS patients recovery of cellular immune function, i.e., it delayed the recovery by about 6 days. CONCLUSION: With or without treatment with glucocorticoids,the lowered CD3+, CD4+, CD8+ T-lymphocyte counts in the early stage are of very important significance in differential diagnosis between severe acute respiratory syndrome and common atypical pneumonia.

Adult↗
PubMed1996

Lymphocyte subset counts during the course of community-acquired pneumonia: evolution according to age, human immunodeficiency virus status, and etiologic microorganisms.

We investigated the evolution of lymphocyte subsets in blood samples from 94 consecutive patients with community-acquired pneumonia; these samples were obtained 72 hours after hospitalization and at follow-up, when the patients were considered to be cured. Acute-phase lymphocyte counts, as compared with follow-up counts, were decreased in the overall population (P < .0001); however, the percentage of CD4+ T cells remained unchanged during the course of pneumonia in each patient. These changes that were noted for the overall population were not significantly influenced by age, the presence (or not) of infection with human immunodeficiency virus (HIV), or etiologic microorganisms. Of the 68 HIV-seronegative patients, 12 had initial CD4+ T cell counts of < 200/mm3; multivariate analysis revealed that this finding was significantly associated with the presence of an underlying disease or tuberculosis and an age of > or = 60 years.

AIDS-Related Opportunistic Infections↗
PubMed1993

Effects of adoptive immunotherapy with autologous CD8+ T lymphocytes on immunologic parameters: lymphocyte subsets and cytotoxic activity.

CD8+ cytotoxic T lymphocytes (CTL) may be an important parameter of host resistance to HIV infection. The present study determined whether CD8+ cells could be purified and propagated in vitro to enhance anti-HIV CTL activity, and the immunologic effects of infusion of these cells into autologous, HIV-infected patients as a potential immunotherapy for AIDS and AIDS-related complex (ARC). CD8+ lymphocytes from five AIDS and ARC patients were purified from leukapheresis preparations in cell culture flasks coated with CD8-specific monoclonal antibodies and propagated in vitro for 3 weeks. The ex vivo propagated cells were 98% (+/- 1%) CD8+ and 43% (+/- 6%) HLA-DR+. The majority of the CD8+ cell preparations had increased lytic activity against autologous B lymphoblastoid cells infected with vaccinia virus vectors expressing HIV-IIIb structural proteins gag, pol, or env, relative to that of fresh blood mononuclear cells tested prior to purification and culture. The results also show for the first time that CD8+ CTL from HIV-infected patients can lyse cells expressing the HIV regulatory protein, tat. Enhanced expression of CD56 (natural killer cell marker) and lytic activity against vaccinia virus control vector-infected, autologous targets were also noted in the CD8+ cell preparations. Infusion of the CD8+ CTL into autologous patients was well-tolerated and resulted in low but discernible, temporal increases in circulating cytotoxic activity against the HIV gene-expressing targets.

AIDS-Related Complex↗
PubMed1994

Effect of long-term administration of beta-carotene on lymphocyte subsets in humans.

To examine whether or not beta-carotene alters immune indexes, we performed a single-blind trial in healthy male nonsmokers. Lymphocyte subsets were assessed by double labeling with monoclonal antibodies before and after 44 wk of administration of either beta-carotene (60 mg/d; n = 10) or placebo (n = 10). In addition, we measured the beta-carotene concentrations in plasma, mononuclear cells (MNC), platelets, and red blood cells (RBCs), as well as the plasma alpha-tocopherol concentration. An increase of plasma and MNC beta-carotene concentrations to four and three times the baseline value was noted after 2 and 4 wk, respectively. The CD4-CD8 ratio increased after 9 mo of beta-carotene administration whereas natural killer cells, virgin T cells, memory T cells, and cytotoxic T cells remained unaltered throughout the study. No side effects or toxicities were detected by a questionnaire survey and routine laboratory tests.

Adult↗
PubMed1991

Periodontal disease in HIV-infected patients in relation to lymphocyte subsets and specific micro-organisms.

Visible plaque index (VPI), gingival bleeding index (GBI) and pocket depth (PD) were analyzed in relation to potential periodontal pathogenic microorganisms and peripheral numbers of T4+ and T8+ lymphocyte subsets in 10 patients with human immunodeficiency virus (HIV) infection, 10 patients with AIDS related complex (ARC) and 10 patients with acquired immune deficiency syndrome (AIDS). 10 healthy persons served as controls. Periodontal disease in patients with more advanced stages of HIV infection were related to the severity of the systemic disease, and to decreasing numbers of T4+ lymphocytes in peripheral blood, but not to VPI or the occurrence of periodontal pathogenic micro-organisms.

AIDS-Related Complex↗
PubMed1987

Quantitation and estimation of lymphocyte subsets in tissue sections. Comparison with flow cytometry.

A quantitation method for lymphocyte subsets in immunoperoxidase-stained frozen tissue sections was compared with flow cytometry in 23 cases of non-Hodgkin's lymphoma. Close correlations were obtained, demonstrating the accuracy of the technic. Weak intensity of fluorescence and fragility of the tumor cells during the fluorescence-activated cell sorter (FACS) analyses were the most likely explanations for a number of the discrepancies observed. The tissue quantitation method was precise, particularly at low values, where it was better than the FACS. A simpler and faster estimation method employing categories within 10 percentage units was also tested in this study; this method correlated as well with the FACS as the quantitation method and gave the best interobserver correlations.

Antigens, Differentiation, T-Lymphocyte↗