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PubMed2002

Spontaneous and induced genetic damage in T lymphocyte subsets evaluated by the Comet assay.

High inter- and intra-individual variability was reported in the level of DNA damage, both spontaneous and induced, when peripheral blood mononuclear leukocytes were used to perform the Comet assay. In order to find out the underlying causes for such variability, different subsets of T lymphocytes were isolated by immunomagnetic cell sorting. The level of DNA damage was evaluated with the alkaline version of the Comet assay by using three different parameters: tail moment, tail length and amount of DNA in the tail (%). Helper T cells (CD4+), cytotoxic T cells (CD8+), their negative fraction and the mixed cell population were evaluated both in untreated cells and after 10 and 20 microM H(2)O(2) treatments. Differences between cell subsets were only observed after H(2)O(2) treatment. The results indicate that, although CD4+ is the fraction with the highest induced level of genetic damage, this value is not high enough to explain the large inter- and intra-individual variability found.

Adult↗
PubMed1997

A case-control study on lymphocytic subsets in elderly bearing a gastroenteric cancer.

The aim of this work is to evaluate the differences in lymphocytic sub-classes between elderly patients with gastroenteric cancer and elderly patients with a non neoplastic disease. A group of 88 patients over 60, consecutively admitted to the III Division General Surgery for gastro-enteric cancer has been collected for the study, the control group consisted of 74 patients also over 60, consecutively admitted over the same period for benign abdominal diseases. In all patients the following data were measured: body mass index (BMI), white blood cells (WBC), total lymphocytes, total T lymphocytes (CD3+), helper T lymphocytes (CD4+), suppressor T lymphocytes (CD8+), CD4+/CD8+ ratio, B lymphocytes, CD5+ B lymphocytes, activated T lymphocytes (CD3+ HLA-DR+), CD4+ "naive" lymphocytes (CD4+ CD45 RA+), CD4+ "memory" lymphocytes (CD4+ CD45 RO+), NK lymphocytes (CD16+ 56+), red blood cells (RBC), total serum cholesterol, albumin, total serum proteins. The main lymphocytic subsets were on an average lower in the cancerous elderly group with respect to the non cancerous. As the tumour progressively increases in size (T), total lymphocytes significantly decrease, while CD4+ progressively decreases with nodal involvement (N). In the cancerous elderly, we found a lower immune response. The immune system appears to be less efficient also in association with tumor growth, especially when T and N get worse. The response of effector cells to the tumour seems not specific.

Aged↗
PubMed1983

Lymphocyte subsets in normal and pre-eclamptic pregnancies.

Peripheral blood lymphocyte subsets in normal and preeclamptic pregnancies have been studied by using the monoclonal antibodies OKT3 (T cells), OKT4 (helper/inducer T cells) and OKT8 (suppressor/cytotoxic T cells). In addition the numbers of mononuclear cells bearing Ia and monocyte antigens have been assessed by using the monoclonal antibodies OKIa1 and OKM1. No significant differences were found between 10 normal pregnant and 10 non-pregnant subjects. Ten preeclamptic patients were studied and showed an increase in OKT4-positive helper cells. This was significant in terms of percentage of mononuclear cells but not the absolute numbers or the helper/suppressor (OKT4/OKT8) ratio.

Adolescent↗
PubMed2004

Immune recovery of lymphocyte subsets 6 years after autologous peripheral blood stem cell transplantation (PBSCT) for lymphoproliferative diseases. A comparison between NHL, HD and MM in group of 149 patients.

To evaluate the normalization of lymphocyte subsets several years after autologous peripheral blood stem cell transplantation (aPBSCT) and to detect any differences based on the underlying lymphoproliferative diseases, we analyzed the immunological recovery of 149 patients with Non Hodgkin's Lymphoma (NHL), Hodgkin's Disease (HD), Multiple Myeloma (MM). Lymphocyte recovery was assessed before the transplant, on days 15, 30, 60, 90, 120 and on years 1, 2, 4, 6. Analysis of a total of 709 lymphocytes, including total lymphocyte count, CD3 +, CD4 +, CD8 +, CD4 +/CD8 + ratio, CD19 +, CD3 + HLA-DR +, CD16 + 56 +, was performed. The normalization of total lymphocyte counts was achieved between days 14 to 22 following PBSCT. CD3 + cells count showed a normalization after 2 years in the HD and NHL groups and after 4 years in MM group. CD4 + subset achieved normalization during the sixth year in the 3 groups. The CD8 + and CD19 + lymphocytes subsets achieved normal values in the 3 groups at day 60 and at day 120 respectively. CD16 + 56 + and CD3 +/HLA-DR + lymphocytes showed median values above the normal range starting from day 30. Immunological recovery was similar in all 3 groups. Moreover, the recovery of all subsets evaluated was similarly demonstrated within 6 years after aPBSCT.

Adolescent↗
PubMed1984

Impaired distribution of T-lymphocyte subsets in patients with haemophilia.

The distribution of T-lymphocyte subsets was assessed using monoclonal antibodies in 20 symptom-free patients with haemophilia. In 15 patients, the T-cell subsets appear substantially balanced with only a moderate reduction in the proportion and absolute number of OKT4 positive cells. In the remaining 5, all with a reversed OKT4/OKT8 ratio, the proportion and absolute number of OKT4 positive cells was significantly reduced (p less than 0.0002) while the absolute number of OKT8 positive cells was normal or reduced although the proportion of these cells appeared increased. Functional studies, testing the proliferative response to PHA and PWM, were normal in all cases including those with immunological abnormalities. These results suggest that a proportion of patients with classic haemophilia show some immunological abnormalities similar to those observed in patients with acquired immunodeficiency syndrome and that regular evaluation of several immunological parameters is warranted in these patients.

Adolescent↗
PubMed1988

[Marginal periodontitis and the immune system. III. Differences in peripheral blood lymphocyte subsets before and after treatment in adult periodontitis patients].

The lymphocyte subsets in the peripheral blood of 8 individuals, aged 25 to 57 yr, with moderate to advanced adult periodontitis were analyzed before and after treatment by means of flow cytometry, and then compared with the findings in 20 normal adults (aged 30 to 45 yr). The results were as follows: the percentage of OKT-8+ (CD 8) in the patients (20.25 +/- 7.15%) was lower than that in normal individuals (31.50 +/- 6.03%, P less than 0.01) while the OKT-4/OKT-8 ratio (2.34 +/- 1.18) was higher than the corresponding ratio in the controls (1.23 +/- 0.37); however, there was no significant difference in the percentages of OKT-3+ (CD 3), OKT-4+ (CD 4) and Leu-12+ (CD 19) between patients and normal subjects. The percentages of OKT-3+, OKT-4+, OKT-8+, Leu-12+ and OKT-4/OKT-8 after the periodontal therapy did not show a significant difference compared with the data obtained before treatment. These results indicate that the decrease in suppressor/cytotoxic T-cell population may be a kind of pathological disposition of the initiation and/or course of adult periodontitis.

Adult↗
PubMed1981

Human B lymphocyte subsets. I. IgG-bearing B cell response to pokeweed mitogen.

The subset of B lymphocytes having IgG on their surfaces was purified from human spleen and blood using a fluorescence-activated cell sorter (FACS). This subset constituted about 15% of B lymphocytes. The remaining non-IgG-bearing B cells were also obtained for study. These two populations were examined for (a) their expression of other surface immunoglobulin isotypes, (b) their ability to give rise to IgG- and IgM-secreting (plaque-forming) cells in a pokeweed mitogen (PWM)-driven culture system, and (c) their ability to proliferate in response to PWM stimulation. The results of these studies indicate that most IgG-bearing B cells also express surface IgM and IgD. Less than 15% had only IgG. The IgG-positive cell gave rise to both IgG and IgM plaque-forming cells when driven by PWM, and in fact were responsible for most of the total plaque response in both the IgG and IgM classes. The non-IgG-bearing B cells were depleted of both IgG and IgM responsiveness. The failure of the non-IgG-bearing B cells to give a strong response to PWM did not appear to be due to either depletion of accessory cells or to any suppressive influence. Finally, proliferation studies indicated that both the IgG-bearing and the non-IgG-bearing cells proliferated in the presence of PWM with a somewhat stronger proliferative response in the IgG-bearing cells. These results demonstrate that the IgG-bearing cell is not irreversibly committed to IgG production but can also give rise to IgM-secreting cells, and that human PWM-driven immunoglobulin secretory responses are predominantly due to a numerically small subset of B cells.

B-Lymphocytes↗
PubMed1997

Study of T-lymphocyte subsets of healthy and Mycobacterium avium subsp. paratuberculosis-infected cattle.

The relative contributions of T-lymphocyte subsets to host defense in cattle infected with Mycobacterium avium subsp. paratuberculosis is reported. The subsets were purified with appropriate monoclonal antibodies and a magnetic bead column separation system, and their purity was verified by flow cytometry. Biological activity of each subset, expressed as lymphoproliferation and gamma interferon (IFN-gamma) production, was measured in response to phytohemagglutinin (PHA) and an M. avium antigen preparation (A-PPD). IFN-gamma was measured by antibody capture enzyme-linked immunosorbent assay. The results showed a correlation between proliferation and IFN-gamma production in response to A-PPD but not to PHA. In response to PHA, CD4+ lymphocytes were the most prolific producers of IFN-gamma. CD8+ lymphocytes produced IFN-gamma to a lesser extent, whereas gammadelta+ T lymphocytes produced little or no IFN-gamma. Differences observed between the amount of IFN-gamma produced by CD4+ versus CD8+ cells and CD4+ versus gammadelta+ cells were significant (P < 0.01), but those between peripheral blood mononuclear cells (PBMC) and CD4+ T cells were not. Similar responses to A-PPD were observed except that PBMC produced higher levels of IFN-gamma than did CD4+ T cells. These data for cattle are similar to observations made for other animal species, where CD4+ cells are the major type of T lymphocytes producing IFN-gamma. They further suggest that whatever the role gammadelta+ T cells may play in paratuberculosis, it is not likely to be mediated by IFN-gamma production.

Animals↗
PubMed1992

Measuring percent lymphocytes by flow cytometry to calculate absolute lymphocyte subset counts for HIV+ specimens.

A comparison was made of lymphocyte percentages from an automated hematology analyzer (ELT 15) vs. a fluorescence flow cytometer (Cytofluorograf). The hematology values were consistently higher than the flow (by greater than 10% for 13 of 50 HIV+ specimens). The findings were similar to three other pairings: H*1 vs. Cytofluorograf, Cell Dyn 3000 vs. Cytofluorograf, and Cell Dyn 3000 vs. EPICS Profile. In another comparison manual percent lymphocytes matched much better with flow values. Factors of sample preparation and instrument analysis as they relate to nonrandom cell loss and lymphocyte resolution are examined.

Acquired Immunodeficiency Syndrome↗
PubMed1983

Effect of lymphoblastoid interferon on lymphocyte subsets in cancer patients.

The effect of interferon dose on lymphocyte subsets is unclear for humans, especially with prolonged intermittent administration. Enumeration of T4 and T8 cells and natural killer (NK) cell activity against K562 cells were examined as part of our ongoing randomized clinical study comparing low- and high-dose treatment strategies. Fifty-four patients have been entered into study, of which 46 are now evaluable. High-dose patients showed no significant changes during actual interferon infusion. However, over the entire period on study, there was a significant increase in NK cell activity (p = 0.001), a concurrent decrease in T8 (p = 0.004), and an increase in T4/T8 (p = 0.02). By contrast, for low-dose patients there was a trend of increasing NK activity during interferon administration, but no change during the total time on study, with a concurrent trend of decreasing T4/T8 and a significant increase in T8 (p = 0.01). Although there is as yet no significant response difference between high- and low-dose treatments, a favourable response was associated with an overall increase in NK activity (p = 0.005), a decrease in T8 (p = 0.02), and an increase in T4/T8 (p = 0.03). In a preliminary study of immune complexes in selected patients, an inverse relationship between NK cell activity and immune complex concentration was suggested. This was supported by in vitro addition of autologous immune complex-containing sera during NK cell assay.

Breast Neoplasms↗
PubMed1984

Autologous mixed lymphocyte reaction in man. IX. Autologous mixed lymphocyte reaction and lymphocyte subsets in aging humans.

Peripheral blood from 15 young (20-30 years) and 15 aging (65-80 years) subjects was analysed for the proliferative response of T cells upon stimulation with non-T cells in the autologous mixed culture reaction (AMLR) and allogeneic MLR, and for the proportion of monoclonal antibody-defined lymphocyte subsets and monocytes. No significant difference was observed in the AMLR or allogeneic MLR between T and non-T cells in aging and young subjects. However, when non-T cells were further fractionated into adherent monocytes and non-adherent B cells (B cells and null cells), the AMLR between macrophages and T cells was significantly (P less than 0.05) higher in young subjects than in simultaneously studied aging subjects. In contrast, the AMLR between T cells and B cells was significantly (P less than 0.05) higher in the aging subjects than in the young group. In the T-T AMLR (using phytohaemagglutinin-stimulated T cells as stimulators), aging subjects had a significantly (P less than 0.05) lower proliferative response than simultaneously studied young subjects. In vitro addition of purified interleukin 2 reconstituted the T-T AMLR to the base-line T-T AMLR in young humans. No significant difference was observed in the allogeneic MLR and lymphocyte subsets between the two groups. The significance of these observations is discussed.

Adult↗
PubMed2005

Effect of chronic ethanol feeding on 24-hour rhythms of mitogenic responses and lymphocyte subset populations in thymus and spleen of peripubertal male rats.

This work analyzes the effect of chronic ethanol feeding on the 24-hour variation of mitogenic responses and lymphocyte subset populations in thymus and spleen. Animals were maintained under a 12:12-hour light/dark photoperiod and they received a liquid diet for 4 weeks, starting on day 35 of life. The ethanol-fed group received a similar diet to controls except that maltose was isocalorically replaced by ethanol. Ethanol replacement provided 36% of the total caloric content of the diet. Rats were killed at 6 time intervals around the clock, beginning at Zeitgeber time (ZT) 1 (ZT 0 = lights on). Under ethanol intake the splenic and thymic weight decreased. In addition, mean values of the thymic, but not of the splenic T cell number decreased, and mean values of the thymic and splenic CD8+ and CD4+CD8+ number increased. Consequently, the thymic T/B ratio and the thymic and splenic CD4+/CD8+ ratio decreased in ethanol-fed rats. At the same time there was a significant increase in the response of the thymic cells to LPS. The ethanol diet modified the 24-hour rhythmicity of thymic and splenic T, B and CD4+CD8+ cells, thymic CD4+ and splenic CD8+ cells, thymic and splenic T/B and CD4+/CD8+ ratios, as well as of mitogenic responses in both tissues. Chronic ethanol administration presumably affects the endogenous clock that modulates the circadian variation of immune responsiveness in growing rats.

Animals↗
PubMed2004

Normal values for CD4 and CD8 lymphocyte subsets in healthy Chinese adults from Shanghai.

The aim of this study was to establish reference ranges for lymphocyte subsets in Chinese adults. Venous blood specimens were obtained from 614 healthy, human immunodeficiency virus (HIV)-seronegative adults in Shanghai. Flow cytometry was used to determine percentages and absolute numbers of CD4 and CD8 T lymphocytes. Mean values for CD4 and CD8 lymphocytes were 727 and 540 cells/microl, respectively, yielding a CD4/CD8 ratio of 1.49. While CD8 lymphocyte values varied with age and gender, no significant differences in CD4 lymphocyte values were observed. Shanghai adults had approximately 100 fewer CD4 lymphocytes/microl on average than Caucasians, suggesting that lower CD4 lymphocyte cutoffs for classifying and monitoring HIV infection may be needed in China.

Adolescent↗
PubMed1991

Natural killer cells and lymphocyte subsets in active MS and acute inflammation of the CNS.

Cerebrospinal fluid (CSF) and peripheral blood (PB) lymphocyte subsets were determined by flow cytometry (FCM) in 15 patients with active multiple sclerosis (MS) and 15 patients with acute inflammatory diseases (ID) of the central nervous system (CNS) in order to establish correlations between the two groups of diseases, as well as between the CSF and PB subsets distribution. A panel of monoclonal antibodies was applied to all the samples: Leu3 (CD4), Leu4 (CD3), Leu2 (CD8), Anti-HLA-DR, Leu11 (CD16). Statistical analysis did not show differences in CD3+ nor in CD3+ DR+ T-cells both in the CSF and PB in the two groups of patients. CD4+ cells were significantly higher in the CSF than in the PB, while CD8+, DR+ CD3- and CD16+ cells were constantly lower in the CSF without differences between the two groups of diseases.

Adult↗
PubMed1988

Blood transit and recirculation kinetics of lymphocyte subsets in normal rats.

Thoracic duct lymphocytes (TDL) were labelled with fluorescein isothiocyanate (FITC) and injected into normal, unanaesthetized rats with either a central venous catheter or a thoracic duct cannula. The blood transit time and the appearance in the lymph was calculated and then the percentages of B, T, T helper, and T suppressor lymphocytes were determined with monoclonal antibodies (Ox12, Ox19, W3/25, and Ox8, respectively). The blood transit time of all subsets was about 30 min. However, the percentage of B lymphocytes from 5 min after injection onwards is reduced (14.5 +/- 1.9%) compared to the injected TDL (32.7 +/- 1.7%). These cells are not in the lung vascular pool. The recovery of FITC-labelled TDL in the thoracic duct within 48 h is much higher (56.8 +/- 5.3% of the injected lymphocytes) than in previous studies using radioactive markers. B lymphocytes appear later and in a reduced number in the thoracic duct. The mean transit time is 26 h and the recovery 31.5 +/- 4.2% in contrast to T lymphocytes (18 h and 66.3 +/- 6.5%, respectively). The technique of combining FITC in vitro labelling with surface staining after completion of migration does not interfere with lymphocyte migration. It can therefore be used to study the migration of lymphocyte subsets in normal, untreated animals.

Animals↗
PubMed2004

Tumour T-lymphocyte subset infiltration and tumour recurrence following curative resection for colorectal cancer.

AIM: The relationship of tumour T-lymphocytic subset infiltration and recurrence in patients undergoing potentially curative resection for colorectal cancer has not been clearly defined. METHODS: Tumour sections from patients who had undergone potentially curative resection for colorectal cancer were stained and counted for CD4+ and CD8+ T-lymphocytes. RESULTS: Twenty-three patients developed recurrence during the follow-up period. Patients were grouped according to whether or not they developed recurrence. The groups were similar in age, sex, site of tumour, Dukes stage and the numbers of patients receiving adjuvant therapy. The total percentage volume of labelled CD4+ T-lymphocytes in the tumour was significantly lower in the patients who recurred (p<0.05). CONCLUSIONS: The results of the present pilot study suggest that a reduction in tumour T-lymphocyte infiltration, in particular CD4+ T-lymphocyte infiltration, is associated with recurrence in patients following potentially curative resection for colorectal cancer.

Aged↗
PubMed1993

Peripheral blood lymphocyte subsets in infants with diarrhea with and without Giardia lamblia infection.

The aim of the present study was to define the cellular immune response during gastrointestinal Giardia lamblia infection in young children. The level of lymphocyte subsets was determined in the peripheral blood of infants with G. lamblia-associated diarrhea or acute gastroenteritis and from control infants without diarrhea. The proportion of peripheral blood lymphocytes (PBL) expressing the CD8 marker (suppressor cytotoxic T cells) and the CD57 marker (natural killer cells and subset of CD8+ T cells) was highest in infants with acute gastroenteritis, lower in infants without diarrhea, and lowest among those with G. lamblia-associated diarrhea. The level of CD4+ PBL (helper T cells) did not differ significantly among the three groups of children tested. The level of memory, or helper-inducer, CD4+CD29+ PBL was increased markedly in acute gastroenteritis as compared with their level among the other two groups, whereas naive or virgin CD4+CD45RA+ PBL had the reciprocal distribution among the three groups of infants. In contrast to acute gastroenteritis from other causes, G. lamblia-associated diarrhea did not elicit changes in lymphocyte subsets.

Antibodies, Monoclonal↗
PubMed1985

[Changes of peripheral lymphocyte subsets in patients with oral squamous carcinoma].

The distribution of peripheral lymphocyte subsets of 32 patients, who had undergone surgical removal of squamous carcinoma and have evaded recurrence was studied using OK series monoclonal antibodies. The T-lymphocyte subpopulation was reduced in patients who were tumor free for more than 3 years. The helper/inducer T-cell subpopulation was reduced in most of the patients studied, whereas little change was observed in the subpopulation of suppressor/cytotoxic T cells. Consequently the ratio of OKT 4+/OKT 8+ was reduced in accordance with the tumor free period.

Adult↗