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PubMed1990

Flow-cytometric analysis of lymphocyte subsets in patients with advanced periodontitis.

Peripheral blood lymphocytes from 36 patients with advanced periodontitis and from 34 healthy subjects were examined using a panel of monoclonal antibodies and fluorescence flow cytometry. The absolute and relative counts of B-cells, T-cells, T-helper (TH), T-suppressor/cytotoxic (TS) cells, activated T-cells and natural killer cells were assessed and the TH/S ratio was calculated. In the periodontitis patients, the TH/S ratio was 1.56 +/- 0.62 and in the controls 1.35 +/- 0.55. This was not statistically significant. A classification on the basis of sex also did not show significant differences in the TH/S ratio. B-cells and activated T-cells were slightly increased in the periodontitis group. However, all determined lymphocyte subpopulations revealed no significant differences between the groups. These results indicate that local inflammatory reactions and immunoregulatory dysfunction are limited to the periodontium in the patients with advanced periodontitis, with no significant quantitative effects on the peripheral lymphocyte subpopulations.

Adult↗
PubMed2005

Lymphocyte subset analyses in healthy adults vaccinated with yellow fever 17DD virus.

In this study the kinetics of humoral and cellular immune responses in first-time vaccinees and re-vaccinees with the yellow fever 17DD vaccine virus was analyzed. Flow cytometric analyses were used to determine percentual values of T and B cells in parallel to the yellow fever neutralizing antibody production. All lymphocyte subsets analyzed were augmented around the 30th post vaccination day, both for first-time vaccinees and re-vaccinees. CD3+ T cells increased from 30.8% (SE +/- 4%) to 61.15% (SE +/- 4.2%), CD4+ T cells from 22.4% (SE +/- 3.6%) to 39.17% (SE +/- 2%) with 43% of these cells corresponding to CD4+CD45RO+ T cells, CD8+ T cells from 15.2% (SE +/- 2.9%) to 27% (SE +/- 3%) with 70% corresponding to CD8+CD45RO+ T cells in first-time vaccinees. In re-vaccinees, the CD3+ T cells increased from 50.7% (SE +/- 3%) to 80% (SE +/- 2.3%), CD4+ T cells from 24.9% (SE +/- 1.4%) to 40% (SE +/- 3%) presenting a percentage of 95% CD4+CD45RO+ T cells, CD8+ T cells from 19.7% (SE +/- 1.8%) to 25% (SE +/- 2%). Among CD8+CD38+ T cells there could be observed an increase from 15 to 41.6% in first-time vaccinees and 20.7 to 62.6% in re-vaccinees. Regarding neutralizing antibodies, the re-vaccinees presented high titers even before re-vaccination. The levels of neutralizing antibodies of first-time vaccinees were similar to those presented by re-vaccinees at day 30 after vaccination, indicating the success of primary vaccination. Our data provide a basis for further studies on immunological behavior of the YF 17DD vaccine.

Adolescent↗
PubMed1988

Analysis of ovine peripheral blood lymphocyte subsets following Ficoll-Hypaque separation or erythrocyte lysis.

Peripheral blood lymphocyte subsets were analysed with the aid of monoclonal antibodies and flow microfluorometry following separation by either the Ficoll-Hypaque method or erythrocyte lysis of the buffy coat. The percentage of lymphocytes labelled by the monoclonal antibodies SBU-T1, SBU-T4 and SBU-T8 were significantly greater (P less than 0.05) in samples obtained by erythrocyte lysis. In contrast, a lower percentage of lymphocytes was labelled by SBU-T6 following erythrocyte lysis (P less than 0.02). These data suggest caution when choosing a method for separation of peripheral blood lymphocytes.

Animals↗
PubMed1989

In situ localization of T lymphocyte subsets in human paracoccidioidomycosis.

Immunohistochemical techniques using monoclonal antibodies to T lymphocyte subsets were used to characterize granulomas caused by Paracoccidioides brasiliensis. Punch biopsies of skin or mucosa from eight patients and of lymph nodes from two patients with disseminated paracoccidioidomycosis were studied. The T lymphocytes were distributed either in a localized pattern related to epithelioid granulomas or in a diffuse arrangement, predominantly around the vessels. In the granulomas, T cells formed a peripheral mantle surrounding central aggregates of macrophages. The majority of lymphocytes were T-helper cells with few suppressor cells. In contrast, patients presented with a decreased number of peripheral T-helper lymphocytes and a corresponding decrease in helper-suppressor cell ratios. There was no clear-cut relationship between tissue helper-suppressor cell ratios and the level of cellular immunodepression of the patients. The lowest P. brasiliensis antibody titers were detected in patients with the highest tissue helper-suppressor cell ratios. The distribution pattern of T lymphocytes in P. brasiliensis granulomas, with a predominance of helper phenotype, suggests that these cells are actively involved in the disease process.

Adolescent↗
PubMed1992

CD4 lymphocyte subset abnormalities associated with impaired delayed cutaneous hypersensitivity reactions in patients with X-linked agammaglobulinaemia.

Circulating CD4 lymphocyte subset (CD45RA; CD45RO; CD29; Leu8) levels were determined in nine patients with X-linked agammaglobulinaemia (XLA), nine patients with common variable immunodeficiency (CVI) and in 18 age- and sex-matched controls. CD4CD45RO and CD4CD29 cells were significantly lower (P less than 0.01) in the XLA patient group (CD45RO, 15.7 +/- 10.2%; CD4CD29, 32.1 +/- 14.6%) compared with CVI patients (61.8 +/- 25.4%; 60.1 +/- 11.2%) and normal controls (43.7 +/- 22.3%, 54.5 +/- 22.0%). The levels of CD4CD45RA and CD4Leu8 cells were not abnormal in the XLA patient group. No selective reduction in CD4 subsets was observed in the CVI patient group. Delayed cutaneous hypersensitivity testing of five XLA and five CVI patients revealed a significantly reduced response to recall antigens in patients with XLA. This may relate to the deficiency of circulating memory T cells observed in these patients.

Adolescent↗
PubMed1982

Enumeration of T-lymphocytes and T-lymphocyte subsets in rheumatoid arthritis using monoclonal antibodies.

The number of T-lymphocytes and T-lymphocyte subsets was measured in peripheral blood of 51 patients with rheumatoid arthritis. T-lymphocytes were counted by E-rosette tests and by the immunogold staining method with OKT3.PAN monoclonal antibody. Helper and suppressor T-lymphocytes were determined by the immunogold staining method with OKT4.IND and OKT8.SUP monoclonal antibody. The relative and absolute numbers of T-lymphocytes and helper T-lymphocytes in peripheral blood of patients with RA did not differ significantly from those in the blood of healthy subjects. However, the relative and absolute numbers of suppressor T-cells were significantly lower in patients with RA than in healthy subjects. The decrease of suppressor T-cells in the blood of patients with RA dit not correlate with the activity of the disease nor the presence of the rheumatoid factor.

Adult↗
PubMed2001

[Study on the distribution of lymphocyte subsets in the eutopic and ectopic endometrium of women with endometriosis].

OBJECTIVE: To investigate the abnormal distribution of lymphocytes in eutopic and ectopic endometrium of patients with endometriosis and its significance. METHODS: In 43 cases, biopsies of ectopic tissues were taken by laparoscopy and laparotomy from patients with endometriosis and eutopic endometrium by curettage at the same time. In 19 cases, eutopic endometrium was taken from hysterectomy for myomatous uterus. Immunohistochemical techniques were employed to demonstrate the difference in the number and ratio of the lymphocyte subsets between the patients with endometriosis and the controls. RESULTS: In the patients with endometriosis, in the proliferative phase, ectopic endometrium contained respectively CD3+ CD8+ T cells and CD68+ macrophages (67.2 +/- 13.5)/5 HP, (45.0 +/- 14.0)/5 HP and (37.2 +/- 10.6)/5 HP, significantly higher then that in the eutopic endometrium (52.4 +/- 11.3)/5 HP (P < 0.01), (32.5 +/- 10.0)/5 HP (P < 0.05), and (30.7 +/- 10.3)/5 HP, and also higher as compared with the control group (52.1 +/- 14.9)/5 HP (P < 0.05), (28.9 +/- 12.7)/5 HP (P < 0.01), (26.3 +/- 9.3)/5 HP (P < 0.05); in the secretory phade, CD8+/CD4+, and CD68+ content was respectively 3.5 +/- 1.2, (40.3 +/- 12.2)/5 HP, higher than that in the control group, 3.2 +/- 0.8 (P < 0.05), (28.6 +/- 10.6)/5 HP (P < 0.01). The number of macrophages was also significantly increased. No cyclic changes in the number of lymphocytes in each subpopulation in ectopic endometrium were found. CONCLUSIONS: In the patients with endometriosis, the changes in T lymphocytes and macrophages are mainly on the endometriotic sites. The infiltration of many lymphocytes and macrophages into the ectopic endometrium formed a chronic inflammatory process. The lymphocytes are not able to clear the ectopic endometrium in the late stages of endometrium, on the contrary, they stimulate the further growth of the endometrium.

Adult↗
PubMed1990

Behaviour of lymphocyte subsets in response to immunotherapy with Propionibacterium avidum KP-40 in cancer patients.

In 15 patients the influence of unspecific immunostimulation/immunomodulation was studied. Patients constituting the therapeutical group suffered from colorectal- and gastric carcinoma, respectively, and were preoperatively treated with 10 mg whole cell preparation of immunomodulating Propionibacterium avidum KP-40. This adjuvant immunotherapy resulted in a significant increase (p less than 0.01) of the natural killer (NK)-cell population, however, total leukocyte and lymphocyte count as well as helper- and suppressor T-lymphocyte subsets did not significantly differ form control values.

Adjuvants, Immunologic↗
PubMed1992

Age-related standards for T lymphocyte subsets based on uninfected children born to human immunodeficiency virus 1-infected women. The European Collaborative Study.

The T lymphocyte subsets in the peripheral blood of 459 uninfected children born to white human immunodeficiency virus 1-infected women included in the European Collaborative Study were measured at regular intervals from birth. More than 2400 observations were used to create smooth age-related reference ranges for CD4 and CD8 counts and percentages, CD4:CD8 ratio and absolute lymphocyte count. Standards are presented for children up to 4 years of age. CD4, CD8 and absolute lymphocyte count rose after birth, peaked at around 6 to 9 months of age and then declined toward adult values. CD4 percentage and CD4:CD8 ratio declined steadily from birth onwards. Centile lines for CD4 count and CD4:CD8 ratio converged markedly with age. For the CD4 values, only 3 to 5% of the variation was attributable to differences between the 10 participating centers. These standards allow T lymphocyte abnormalities to be used more effectively as markers for disease progression and assist in the clinical follow up of human immunodeficiency virus 1-infected children. They also provide a basis for initiating antiretroviral treatment or antimicrobial prophylaxis.

Age Factors↗
PubMed1999

Flow cytometry of cerebrospinal fluid (CSF) lymphocytes: alterations of blood/CSF ratios of lymphocyte subsets in inflammation disorders of human central nervous system (CNS).

Flow cytometry was adapted to measure lymphocytes in human cerebrospinal fluid (CSF). The method was sufficiently precise, reproducible and accurate despite low cell counts. In lumbar CSF of controls with 500 to 3500 (10(3)/l) leukocytes, lymphocyte counts correlated with those in corresponding venous blood: blood/CSF ratios of approximately 2000 : 1 were found for total T cells (CD3+) and CD3+ HLA-DR-, CD3+4+, CD3+8+ subsets, ratios were increased for the lymphocyte subsets CD3+ HLA-DR+ < or = CD3+16+56+ < CD16+56+3- < CD8+3- << CD19+; CD8+4+ ratio was half of CD3+ ratio. Data indicate selective barriers (blood-brain and blood-CSF barriers) to blood lymphocyte subsets which favor the transfer of T subsets. Correlation of the subset ratios to the CD3+ ratio indicates distinct barrier properties which changed differently with acute and subacute inflammations and neuroimmunological diseases of central nervous system (CNS) in lumbar or ventricular CSF, but not with simple protein barrier disturbance. HLA DR+ T ratios were higher than HLA DR- T ratios only with controls and some neuroimmunological diseases. Lymphocyte barrier characteristics were related to protein leakage situated at the same barriers, indicating for the lymphocyte subsets selective transfer routes in control subjects and non-selective routes in patients with CNS inflammation where altered ratios revealed a mixture of both routes.

Adult↗
PubMed1998

Peripheral blood lymphocyte subset shifts in patients with untreated hematological tumors: evidence for systemic activation of the T cell compartment.

Flow cytometry immunophenotyping of peripheral blood lymphocyte subsets and multivariate data-analytical techniques revealed that among untreated hemato-oncological patients (n = 48) with lymphomas, acute and chronic myeloid and lymphocytic leukemias, monoclonal gammopathy of undetermined significance, and multiple myeloma, 42% had (nonmalignant) lymphocyte profiles clearly distinct from healthy donors. Notably, a similar pattern of increased CD3+ CD57+, CD3+ HLA-DR+, CD3+ CD(16 + 56)+, CD4- CD8+, CD8+ CD57+, CD8+ CD28-, and CD8+ CD62L- subsets was detected. More extensive three-color immunophenotyping on an additional group of 49 untreated patients revealed that both CD4+ and CD8+ T cells displayed significant increases of activation markers: CD69, CD(16 + 56), HLA-DR, CD71, and CD57, and a loss of CD62L and CD28, which is also interpreted as a sign of activation. Consistent with the phenotypical signs of in vivo immune activation, polyclonal cytolytic activity, measured ex vivo in an anti-CD3-redirected assay, was detected within immunomagnetically purified CD4+ T cells of three out of six B-CLL patients investigated, but not within purified CD4+ T cells of five healthy donors. The purified CD8+ T cells of patients (n = 28) and donors (n = 5) on the other hand displayed similar polyclonal cytotoxic activities at the various effector:target ratios investigated. Tumor-directed cytotoxic activity of purified CD4+ (n = 6) and/or CD8+ T cells (n = 15) against freshly isolated autologous tumor cells was not detected in any of the experiments. Collectively, our results demonstrate systemic T cell activation as a common feature in hematological neoplasia, and a markedly enhanced cytolytic activity of the CD4- subset in CLL patients. The reason(s) for this expansion of activated T cells and its pathophysiologic significance, however, remain unclear.

Adolescent↗
PubMed1996

Gonadotropin-releasing hormone agonist influences absolute levels of lymphocyte subsets in vivo in male mice.

Our earlier studies have demonstrated that gonadotropin-releasing hormone (GnRH) agonists suppress immune system function in female mice. No systematic studies regarding the effect of gender on immune system function following GnRH agonist treatment, however, have been reported. This study, therefore, investigated sequential changes in lymphocyte subsets in 3- and 10-week-old male mice following agonist or placebo administration. Changes in immunophenotypic expression of lymphocytes from thymus, bone marrow, spleen, and blood were analysed at periodic intervals. Upon agonist administration, plasma testosterone levels were significantly increased in pre-pubertal mice, but were significantly decreased in post-pubertal males. Absolute thymic weights, thymocytes and T subsets were significantly increased from the third week regardless of gonadal status. Blood lymphocyte subsets showed a decreasing trend after agonist administration in pre-pubertal males, whereas no differences were observed in post-pubertal males. No significant differences were observed in spleen cells after agonist administration. These studies, together with earlier observations in female mice indicate that GnRH agonist effects on the immune system, are independent of steroid hormone levels. In contrast to suppressive effects in females, GnRH agonist induce no change or ultimately enhanced lymphocyte counts in males, indicating differential effects on the immune system between males and females. This may have important implications for the treatment of various diseases.

Animals↗
PubMed2002

Influence of low-dose oral contraception on peripheral blood lymphocyte subsets at particular phases of the hormonal cycle.

OBJECTIVE: To investigate the effects of low-dose oral hormonal contraception on the immune system during certain phases of the hormonal cycle. DESIGN: Prospective, nonrandomized, controlled study. SETTING: Academic research setting. PATIENT(S): Women with regular menstrual cycle using hormonal oral contraception (OC; Cileste, 250 microg of norgestimat and 35 microg of ethinylestradiol, or Marvelon, 150 microg of desogestrel and 30 microg of ethinylestradiol) and women not using hormonal or other forms of contraception. INTERVENTION(S): Peripheral blood lymphocyte subsets were determined by flow cytometry on the first day of menstruation (day 1), in the follicular phase (day 8), midcycle (day 15), and in the luteal phase (day 22). MAIN OUTCOME MEASURE(S): Levels of lymphocyte subpopulations. RESULT(S): Women using OC had significantly higher levels of CD3+ CD8+ cells throughout their pill cycle compared to controls. Furthermore, women taking Cileste had lower levels of natural killer (NK) cells during their cycle and also women taking Marvelon but only from days 8-15. Within the pill cycle of Cileste we observed an increase in CD20+ and CD20+ CD5+ cells from days 1-8. CONCLUSION(S): Cytotoxic lymphocytes, which are responsible for first-line immune defense, and B cells, which are involved in autoimmune disorders, are affected by OC.

Adult↗
PubMed1985

HNK-1+ (Leu-7) and other lymphocyte subsets in long-term survivors with renal allotransplants.

At 5 or more years after renal transplantation, 42 patients were studied for their lymphocyte subsets to Leu-1, Leu-2, Leu-3, and Leu-7. It was found that, in this group of patients who ranged from 25 to 60 years of age, there was a significant decrease in the number of T helper cells and a decrease in the absolute level of T lymphocytes, with no significant change in the number of T suppressor cells. On a relative basis, the helper/suppressor ratio was decreased in patients when compared with normal persons. This was due to an increase in the relative numbers of suppressor cells. It was demonstrated that the Leu-7+ subset, which marks the NK population, was significantly elevated, in relative proportion, in peripheral blood when compared with controls. This was not seen on an absolute basis. The age-dependence of the relative numbers of Leu-7+ cells was seen in the normal control population and in the transplant cohort. There was no significant correlation between lymphocyte subset measurements and delayed-type hypersensitivity skin tests in the transplant population. A finding of interest is that 6 of the patients who had been treated for malignant disease during their posttransplant course had significantly higher numbers of Leu-7+ cells on a relative basis. In 5 of these patients, for whom data was available on absolute numbers, there was also a highly significant difference in the absolute numbers of Leu-7+ cells in these treated and surviving allograft recipients. It is speculated that this finding may suggest that an increase in Leu-7+ cells marks posttransplant patients who have a successful outcome following the treatment of malignancy, for which they are at increased risk.

Adult↗
PubMed1989

[A study of peripheral blood lymphocyte subsets in patients with bladder tumors].

The analysis of peripheral blood lymphocytic subpopulations was conducted with 16 bladder tumor patients. Lymphocytes were collected centrifugally from the peripheral blood of preoperative patients with bladder tumors, and measured by means of flow cytometry. Among the stages of the bladder tumor patients, there were no clear relationships in the ratio between T cells and B cells. However, with the progression in stages a rise in OKT 8, as well as the lowering of the OKT 4/OKT 8 ratio were discerned. In relation to pTis, Leu 7 showed a higher value than that in other groups with advanced invasion. This suggests that in measurements of lymphocyte subsets in peripheral blood, it will to a certain extent be possible to estimate the preoperative immunological competence of bladder tumor patients.

Humans↗
PubMed1998

Changes in lymphocyte subsets during normal pregnancy.

Peripheral blood lymphocytes from healthy women were studied during pregnancy and postnatally, and were compared with lymphocytes from an age-matched non-pregnant control group. Compared with non-pregnant women, the total white cell count was significantly increased at all pregnancies and also post-partum. In pregnancy the absolute number and percentage of T lymphocytes was slightly elevated while almost no changes in B cells were found. No significant changes were found in the percentage of suppressor/cytotoxic (CD8+), of helper/inducer (CD4+) T lymphocytes, nor of CD4+/CD8+ ratio at any stage of pregnancy and puerperium. The most remarkable changes of the immune system occurred in the group of HLA-DR+ and CD56+ activated T cells. The cell numbers showed a significant increase in the first trimester (< 14 weeks) and decreased slightly from stage to stage. Lower values in NK (natural killer) cells and higher levels of IL-2 receptor positive T lymphocytes did not reach significant levels of change.

Adult↗
PubMed1989

Migration pattern of lymphocyte subsets in the normal rat and the influence of splenic tissue.

Lymphocyte subsets leave the blood and appear in the thoracic duct of normal rats at different rates. The aim of the present study was to investigate their migration pattern through blood, spleen, bone marrow, mesenteric lymph nodes, and Peyer's patches in normal Lewis rats and to study the role of the spleen using splenectomized and spleen-transplanted animals. Fluorescein isothiocyanate (FITC)-labelled thoracic duct lymphocytes (TDL) were injected intravenously into rats and after 15 min, 1, 6, and 24 h the percentages of B, T, T helper (TH) and T-cytotoxic/suppressor (TC/S) lymphocytes in the FITC+ cells were determined in cell suspensions by means of monoclonal antibodies. B and T lymphocytes are preferentially localized in different organs, e.g. B cells in Peyer's patches and T cells in mesenteric lymph nodes. The migration of TH lymphocytes differed from that of TC/S lymphocytes in all the organs investigated. In the late phase after injection the migration of B and TH lymphocytes was influenced by the spleen, since after splenectomy the number of injected B lymphocytes increased and that of TH lymphocytes decreased in all organs investigated except the bone marrow. Splenic autotransplantation could not normalize the disturbed migration.

Animals↗
PubMed1989

[Basic studies on oral administration of lentinan (I)--influence on lymphocyte subsets in peripheral venous blood].

The effect of oral administration of lentinan (LTN), a biological response modifier, in the control of systemic immune function was studied in 6-week old male Wistar-Imamichi SPF rats. In the LTN group, 1 mg LTN dissolved in 1 ml physiological saline was administration forcibly into the stomach twice weekly. Physiological saline alone was administered in a similar fashion to the control group. Blood samples were obtained prior to and after four and eight weeks of administration. White blood cells and lymphocyte counts were obtained and lymphocyte subsets were measured using monoclonal antibodies W3/13, W3/25 and 0 X 8 (Sera-Lab), and a laser flow cytometry system (Orthospectrum III, Orthodiagnostic System). The T cell ratio, helper/inducer T (Th) cell ratio, and suppressor/cytotoxic T (Ts) cell ratio were measured. The peripheral white blood cell count and lymphocyte count were not significantly different between the control and LTN groups. After four weeks of LTN administration, however, the LTN group showed a significantly higher T cell ratio, Th cell ratio and Th/Ts cell ratio than did the control group, and the Ts cell ratio was significantly lower. In the groups undergoing administration for eight weeks, no difference was noted in the lymphocyte subsets between the two groups. Oral administration of LTN apparently modulates the systemic immune function through T cell stimulation, especially Th cells, but continued administration may induce a tolerance to the effect of LTN.

Administration, Oral↗