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Corticosteroid effects on circulating lymphocyte subset levels in normal humans.

The effects of in vivo corticosteroid administration on levels of lymphocyte subsets in normal humans require further definition. Using monoclonal antibodies, we carried out a double-blind, placebo-controlled study of oral and intravenous methylprednisolone in 10 normal volunteers. Four hours after a 7-day oral course of 0.5 mg/kg/day, there was modest lymphopenia but no significant selective alteration in lymphocyte subsets. In contrast, 4 hr after a single intravenous injection of 1.0 mg/kg, there was more pronounced lymphopenia (P less than 0.01), a selective, relative decrease in T4 cells (P less than 0.001), and a more modest decrease in the percentage of T3 cells. The possible mechanisms and implication of these changes are discussed.

Administration, Oral↗

Changes in the host lymphocyte subsets during chemical carcinogenesis.

Changes in small lymphocyte subsets in the lymphoid organs of young C3H mice were studied following i.m. injection of a carcinogenic dose of 3-methylcholanthrene in trioctanoin oil. Using monoclonal anti-Lyt antibodies and a sandwich radiolabeling method with 125I-labeled rabbit anti-mouse Immunoglobulin, the lymphocyte subpopulations in the thymus, spleen, and draining lymph node were examined by radioautography. During the fifth week following the administration of the carcinogen and prior to the appearance of histologically evident tumor cells, a sharp decrease in the level of Ly-1,2+ small lymphocyte population in the thymus was noted which coincided with a considerable increase (10-fold) in the Ly-2+ and a small increase (1.7-fold) in the Ly-1+ population. During the same period, a similar increase in the Ly-2+ population was also observed in the draining but not in the contralateral lymph node. The high levels of Ly-2+ cells lasted for more than 4 weeks in the thymus while, in the draining node, they lasted for 2 weeks and dropped to normal levels (0 to 2%) simultaneously with the appearance of tumor cells identified in histological preparations. Smaller increases (3-fold) in the Ly-2+ subset were also noted in the spleen and contralateral node, but they were seen later (7 to 8 weeks) and were shorter in duration. These systemic increases coincided with the appearance of macroscopic tumor nodules. The relative incidence of immunoglobulin+ cells did not change significantly in any of the organs tested during the entire tumor induction phase, while the level of null cells underwent a slight increase (approximately 2-fold) in all organs tested immediately prior to and following the appearance of macroscopic tumors. None of these changes was observed in trioctanoin oil-injected control animals. The mixed lymphocyte reaction response of the draining node cells, but not of the spleen, was suppressed during the period of increased level of Ly-2+ cells. Furthermore, during this period, s.c. transplantation of a syngeneic mammary tumor in the same leg resulted in enhanced local growth as well as metastatic spread of the tumor to the lungs in 3-methylcholanthrene-treated mice. These findings suggest that a localized immunosuppression associated with the rise in the Ly-2+ cells may be of functional significance during carcinogen-induced tumor development.

Animals↗

T-lymphocyte-subsets in endocrine exophthalmos.

Peripheral T-Lymphocyte subsets were analyzed with monolateral antibodies in 40 patients with Graves' ophthalmopathy. The 20 patients with untreated hyperthyroid exophthalmos showed a slight statistically not significant decrease in the percentage of total T-Lymphocytes and a statistically significant decrease with percentage of the OK T8 cells. No significant changes were observed in the percentage of OK T3 and OK T4 and OK T8 cells in patients with euthyroid exophthalmos under the drug treatment. These findings indicate the significant association of quantitative abnormality of suppressor-cytotoxic cells with untreated hyperthyroid exophthalmos.

Exophthalmos↗

Peripheral blood T lymphocyte subsets in active rheumatoid arthritis--effects of different therapies on previously untreated patients.

Peripheral blood T lymphocyte subset levels were prospectively analyzed in previously untreated patients with active rheumatoid arthritis (RA) with different drug regimens. Before treatment the patient group had reduced numbers of Leu 2a+ lymphocytes compared with controls (p less than 0.001). During penicillamine and chloroquine treatment a marked reduction in absolute numbers of Leu 3a+ lymphocytes was seen (p less than 0.01), whereas treatment with nonsteroidal antiinflammatory drugs was characterized by increased absolute numbers of Leu 2a+ lymphocytes (p less than 0.02). Thus, there seems to be a predictable variation of T lymphocyte subsets in RA with certain drug regimens.

Adolescent↗

Correlation between peripheral lymphocyte subsets and antibody to hepatitis B surface antigen response in hepatitis B vaccine recipients.

Peripheral lymphocyte subsets of 14 heterosexual recipients of hepatitis B vaccine were measured by indirect immunofluorescence using specific monoclonal antibodies. 12/14 vaccine recipients developed antibody to hepatitis B surface antigen after the second injection of vaccine; however, 2 recipients remained seronegative. Prior to the vaccination, these two vaccine non-responders had been found to have a significantly reduced ratio of peripheral helper-inducer T cells to suppressor/cytotoxic T cells (1.15 +/- 0.02 vs. 1.79 +/- 0.12), and an increase in the proportion of natural killer/killer cells (25.6 +/- 1.5% vs. 13.8 +/- 1.6%) as compared to the 12 responders. Enumeration of peripheral lymphocyte subsets prior to vaccination may be useful in predicting the immune response of hepatitis B vaccine recipients.

Adult↗

Evaluation of lymphocyte subsets and NK cells in septic children.

BACKGROUND: Disorders of lymphocyte subpopulations counts and NK cells may not only contribute to impaired elimination of pathogens, but also be an important factor in the development of generalized inflammatory reaction. The aim of the paper was the analysis of subpopulations of lymphocytes and NK cells in children with sepsis and after recovery. MATERIAL/METHODS: The study comprised 25 septic children (17 infants and 8 children aged from 1 to 3 years). The control group consisted of 25 healthy children. The study was conducted in the initial phase of sepsis and then repeated 2 to 3 months after recovery. In all the children, the percentage and absolute numbers of basic lymphocyte subsets and NK cells in blood serum were evaluated. RESULTS: In the course of sepsis, the absolute number of activated lymphocytes T in the youngest children was higher in comparison with the control group. After recovery, the percentage and absolute counts of CD4 lymphocytes in all the children and CD3/CD4 lymphocytes ratio in the youngest children were considerably lower than in the control group. The percentage and absolute number of NK cells in the youngest children after recovery were higher in comparison with the control group. CONCLUSIONS: In children who underwent sepsis, depression traits of cellular immunity are observed. The occurrence of defensive system disorders after recovery may be an important factor in recurrent infections and creates the necessity of immune system monitoring in children with the history of sepsis for the purpose of prevention.

Case-Control Studies↗

[Changes in lymphocyte subsets with post-operative chemotherapy in breast carcinoma--a study of using two color analysis].

Thirty postoperative patients with breast cancer (stage I, II) were studied to clarify the changes of peripheral lymphocyte subsets by administration of anticancer drugs (UFT, cyclophosphamide (CPM]. The patients were divided into two groups: UFT (400 mg/day) group (n = 15) and CPM (100 mg/day) group (n = 15). Immunological investigation, in particular two color analysis of peripheral lymphocyte subsets, were carried out 30 days after treatment. Results from the investigation, there were no inhibitory effects on the total lymphocyte counts by UFT, whereas a promoting effect was observed on Ts (Suppressor T), and CD16+ (NK) subsets. On the other hand, CPM displayed an inhibitory tendency on the total lymphocyte counts, and also an inhibitory effect on Thi (Helper inducer T), and Tsi (Suppressor inducer T) subsets. These results suggest that the changes of lymphocyte subsets in response to above two drugs are different, which may be significant in determining treatment regimens.

Antibodies, Monoclonal↗

Lymphocyte subset differences in patients with chronic fatigue syndrome, multiple sclerosis and major depression.

Chronic fatigue syndrome (CFS) is a heterogeneous disorder of unknown aetiology characterized by debilitating fatigue, along with other symptoms, for at least 6 months. Many studies demonstrate probable involvement of the central and autonomic nervous system, as well as a state of generalized immune activation and selective immune dysfunction in patients with CFS. The aim of this study was to compare the lymphocyte subsets of patients with chronic fatigue syndrome to those of patients with major depression and multiple sclerosis as well as those of healthy control subjects. No differences were found in total numbers of T cells, B cells or natural killer (NK) cells. However, differences were found in T, B and NK cell subsets. Patients with major depression had significantly fewer resting T (CD3(+)/CD25(-)) cells than the other groups. Patients with major depression also had significantly more CD20(+)/CD5(+) B cells, a subset associated with the production of autoantibodies. Compared to patients with multiple sclerosis, patients with CFS had greater numbers of CD16(+)/CD3(-) NK cells. Further study will be required to determine whether these alterations in lymphocyte subsets are directly involved in the pathophysiology of these disorders, or are secondary effects of the causal agent(s).

Adult↗

Discrimination of two different clinical entities, acute-type and subacute-type, human fulminant hepatitis by peripheral blood lymphocyte subsets.

Human fulminant hepatitis tends to be classified into two groups: acute type (A-FH) and subacute type (S-FH). In order to define these two clinical entities more precisely, we examined and compared peripheral blood lymphocyte subsets in A-FH and S-FH patients. We found that S-FH patients had a higher prevalence of CD19+ B cells (31.1 +/- 7.6% in S-FH vs 12.7 +/- 3.7% in A-FH) and also a lower prevalence of CD3+T cells (50.2 +/- 8.7% in S-FH vs 65.6 +/- 10.5% in A-FH). Furthermore, by examining the absolute cell numbers of these subsets, we determined that their imbalance in S-FH was mainly due to a decrease in CD3+ T cells. Among several T cell subsets, the CD8+CD11b-T cell subset was elevated in A-FH and decreased in S-FH (6.1 +/- 2.1% in S-FH, 24.4 +/- 5.8% in A-FH, and 14.8 +/- 7.8% in control). Serial studies of two S-FH patients revealed that the imbalance of these lymphocyte subsets returned to their proper ratio together with the improvement of their liver injury. These results indicate that there might be a different immunological background between A-FH and S-FH.

B-Lymphocyte Subsets↗

Effects of thyroid hormones on the lymphocyte phenotypes in rats: changes in lymphocyte subsets related to thyroid function.

The effects of hyperthyroidism and hypothyroidism on the immune function were assessed in rats by the changes in lymphocyte subsets in peripheral blood and spleen. Hyperthyroidism was induced in rats by intraperitoneal injection of large doses of T4. Hypothyroid rats were prepared by total thyroidectomy followed by 131I administration. In the hyperthyroid rats, the peripheral blood helper/suppressor T cell ratio was decreased, while in hypothyroidism this ratio was increased. However, in spleen cells, this ratio was increased in hypothyroid rats as compared to normal controls. The most striking finding was an augmentation of activated T cells (transferrin receptor positive lymphocytes) in both peripheral blood and spleen cells in the hypothyroid rat. These results suggested that thyroid hormone suppressed the immune response and that a deficiency of this hormone was associated with an increase of activated T cells. These effects of thyroid hormone on the lymphocyte subsets in rats were at variance with the abnormalities that have been observed in humans with such autoimmune thyroid disease as Graves' disease.

Animals↗

A comparative study on the effect of BCG-PSN and thymopeptides on T-lymphocyte subsets of normal and immunosuppressed mice.

To compare the effects of polysaccharide nucleic acid fraction of bacillus calmette guerin (BCG-PSN) and thymopeptides on T-lymphocytes of normal and immunosuppressed mice, CD4+ and CD8+ T-lymphocyte subsets of single nucleic cell in thymus, spleen and peripheral blood were detected successively by flow cytometry after application of BCG-PSN and thymopeptides. Meanwhile, CD4+/CD8+ ratio was also calculated. The results showed that both BCG-PSN and thymopeptides could decrease the proportion of CD4+ CD8+ T-lymphocyte subsets in the thymus, at the same time increase CD4+ T-lymphocyte, CD8+ T-lymphocyte proportion in the three tissues. The fluctuation in amplitude was greater in thymopeptides group than that in BCG-PSN group. It is concluded that acting location of thymopeptides is in thymus, its stimulating action is stronger than that of BCG-PSN, while BCG-PSN not only accelerates the differentiation in thymus, but also has some direct stimulation to peripheral CD4+ T-lymphocytes, and can maintain CD4+/CD8+ ratio within normal range. So, BCG-PSN is safer.

Adjuvants, Immunologic↗

Age-related changes in blood lymphocyte subsets of Saudi Arabian healthy children.

The age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T-lymphocyte population, the CD8(+) subset increased but the CD4(+) subset decreased, resulting in a declining CD4(+)/CD8(+) ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA-DR+ T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease-related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups.

Adolescent↗

Distinct binding patterns of fucose-specific lectins from Biomphalaria alexandrina and Lotus tetragonolobus to murine lymphocyte subsets.

The putative expression of distinct terminally fucosylated glycoconjugates among murine lymphocyte subpopulations was sought using a Biomphalaria alexandrina-derived lectin (BaSI), of proven specificity to a fucosyllactose determinant, and the fucose-binding lectin from Lotus tetragonolobus seeds. Direct labeling of isolated lymphocyte subsets in suspension as well as immuno-histochemical and two-dimensional Western blotting assays demonstrated the exclusive expression of the BaSI-reactive ligand among multiple isoforms of two major 95 and 92 kDa and a minor 82 kDa acidic glycoproteins, selectively localized to the splenic marginal zone B lymphocytes of adult mice. The expression of the L. tetragonolobus lectin-reactive ligand was, on the other hand, restricted primarily to a single homogeneous 50 kDa acidic glycoprotein associated with a subset of the mature (PNA-) medullary thymocytes of adult mice as well as a minority of the immature (PNA +) thymocytes within the deep cortical region in newly born mice. The significance of these findings is discussed in relation to mechanisms that govern lymphocyte development and homing.

Animals↗

T-lymphocyte subsets and eosinophil counts in acute and convalescence chickenpox infection: a household study in Guinea-Bissau.

OBJECTIVE: To investigate changes in T-lymphocyte subsets, CD4+ and CD8+ lymphocytes, WBC, lymphocytes and eosinophil granulocytes during the acute and the convalescence phase of chickenpox infection. METHODS: During an epidemic of chickenpox, a household study was performed in a semi-urban area of Bissau, Guinea-Bissau. Varicella antibodies were determined to assess diagnostic certainty. To determine the timing of changes, haematological markers and T-cell subsets (immunocytochemical method) were analysed in the acute phase, 0-9 days after the rash, and in the convalescence phase, 35-45 days after the rash. RESULTS: In the acute phase, the CD4 percentage, CD4/CD8 ratio, and neutrophil percentage declined, whereas the CD8 percentage, WBC, CD4 and CD8 counts, and the lymphocyte percentage increased over the same period, most markedly for the CD8 count. The eosinophil percentage increased significantly with time from onset of rash. Between acute and convalescence samples there was an increase in CD4 percentage, CD4/CD8 ratio, and CD4 count, and a marked decrease in CD8 percentage and CD8 count. The changes were not significant for WBC, lymphocyte percentage, neutrophil percentage, and monocyte percentage, but eosinophil percentage was significantly increased 5-7 weeks after the onset of rash. The haematological changes were related to number of pox and intensity of exposure; a high eosinophil percentage was associated with less severe disease, i.e. less pox. CONCLUSION: We report significant changes in T-lymphocyte subsets during the acute phase of chickenpox infection, including a suppression of CD4+ T-cells and an augmentation of CD8+ T-cells. The levels were normalized 1 month later except for eosinophils, and we found no persistent CD4 suppression after chickenpox. An increased number of eosinophils in the peripheral blood was demonstrated early in the acute phase of the disease, and remained elevated in the convalescence phase.

Acute Disease↗

Enumeration of peripheral lymphocyte subsets using 6 vs. 4 color staining: a clinical evaluation of a new flowcytometer.

UNLABELLED: Technological advances in instruments allow the evaluation of many lymphocyte subsets in one step. The aim of this study was to evaluate the new FACSCanto flowcytometer in routine conditions, using a 6 color combination, single platform, whole blood, lysis, no wash protocol. METHODS: Two systems were simultaneously compared on 67 blood samples and external quality controls, using CD3,CD4, CD8, CD19, CD16/56, and CD45 in one tube TRUCOUNT beads (BD Biosciences) or two tubes (TetraChrome and Flowcount, Beckman-Coulter and DakoCytomation). RESULTS: The day-to-day instrument detection but automatic compensations were stable. Manual compensation settings were satisfactory using available facilities. Commercial and UK NEQAS quality control results were acceptable. The intra-experiment reproducibility was good (coefficient of variation (CV)<3%) but highly operator-dependent (CD4+ T cell count CVs from 1.2 to 9.7, six operators). Storage of samples was acceptable, but storage of stained samples altered absolute count reliability. Serial dilutions show a good count accuracy. The FACScanto T subsets and B cell data were highly correlated with our reference values (r2>0.87) and absolutes values were very close (slopes>0.89). The gating strategy, fluorochrome choice, and compensation setting are discussed. A few improvements are expected (sample loader, data management, auto-gating, acquisition parameters, sample mixing, absolute values calculation, etc). In conclusions, despite its complexity, 6 color staining is a reliable, stable, and highly informative technique for lymphocyte subset monitoring but remains to be optimized.

Adolescent↗

T-lymphocyte subsets as noninvasive markers of cardiomyopathy.

Fifty-eight patients with symptomatic congestive heart failure were examined for T-lymphocyte subsets in the peripheral blood using two-color laser flow cytometry as a noninvasive diagnostic procedure. The final diagnosis established by catheterization and endomyocardial biopsy were dilated cardiomyopathy (DCM, n = 24), myocarditis (MC) by the Dallas criteria (n = 12), and coronary heart disease (CHD, n = 16). The CD8+CD11- (cytotoxic T) subset was significantly low in patients with DCM (13.9 +/- 4.4 vs. controls, p less than 0.05) in comparison with MC (20.7 +/- 10.9) and CHD (22.3 +/- 5.9). Moreover, the CD4+2H4+ (suppressor/inducer T) subsets were higher in patients with DCM (27.3 +/- 6.9 vs. controls, p less than 0.01) than in those with MC (17.3 +/- 7.8) and CHD (15.6 +/- 7.9). The CD4/CD8 and CD4+2H4+/CD8+CD11- ratio were examined and compared with those of normal controls (NC n = 16). The CD4+2H4+/CD8+CD11- ratio was clearly higher in patients with DCM (2.2 +/- 0.9 vs. controls, p less than 0001) than in those with MC (1.1 +/- 0.6) CHD (0.9 +/- 0.7). A CD4+2H4+/CD8+CD11- ratio of greater than 1.6 was considered to facilitate diagnosis of dilated cardiomyopathy with 79% sensitivity and 70% specificity. There was no significant increase in the ratios between MC and CHD. However, the proportion of the CD8+Leu7+ (natural suppressor) subset of circulating T lymphocytes in patients with MC was statistically higher (19.1 +/- 6.3% vs. controls, p less than 0.05) than in DCM or CHD. An elevated ratio of CD4+2H4+/CD8+CD11- among peripheral blood lymphocytes may thus be a useful marker for differential diagnosis of dilated chronic cardiomyopathy from myocarditis and coronary heart disease.

Adult↗

Diurnal variation of lymphocyte subsets identified by monoclonal antibodies.

Monoclonal antibodies specific for lymphocyte subsets were used to examine circulating lymphocytes obtained at frequent intervals from healthy subjects. A diurnal rhythm was found in the total numbers of lymphocytes, T cells, inducer/helper cells, suppressor/cytotoxic cells, Ia positive cells, and B cells. The lowest levels of all subsets were seen at 0900 hours and the highest levels at 2100. In some subjects the ratio of helper to suppressor cells varied considerably during the sample period, though the ratio was relatively constant for the group as a whole.

Adult↗

Lymphocyte subset counts in continuous ambulatory peritoneal dialysis patients in relation to administration of recombinant human erythropoietin and angiotensin-converting enzyme inhibitors.

Estimation of lymphocyte subset counts (SLCs) is a useful tool in diagnosing nutrition and immune changes in continuous ambulatory peritoneal dialysis (CAPD) patients. Administration of recombinant human erythropoietin (rHuEPO) or angiotensin-converting enzyme inhibitors (ACEIs) can influence SLCs. Our aim was to evaluate the relationship between rHuEPO and ACEI doses and SLCs in the course of CAPD. In the group of studied patients (n = 55), 34 patients were taking rHuEPO and 38 patients were taking ACEIs. In 35 patients, enalapril was the ACEI used. Seven patients were taking rHuEPO, but not ACEIs; 11 patients were taking ACEIs, but not rHuEPO; 27 patients were taking rHuEPO and ACEIs both; and 10 patients were receiving neither rHuEPO nor ACEIs. Flow cytometry was used to estimate CD3, CD4, CD8, CD19, and CD16 + 56 antigens. In the study group, a correlation was seen between dialysis duration and rHuEPO dose (r = 0.395). No correlation was seen between CAPD duration and ACEI dose, but it was seen for total rHuEPO and ACEI doses (r = 0.327). A negative correlation was also seen between dialysis duration and CD19 cell count (r = -0.313). In patients taking only ACEIs (n = 11), a negative correlation was seen between total ACEI doses and CD16 + 56 cell count (r = -0.710). In patients who were not receiving rHuEPO or ACEIs, negative correlations were seen between dialysis duration and total lymphocyte count (r = -0.727), CD3 cell count (r = -0.706), CD4 cell count (r = -0.636), and CD8 cell count (r = -0.764). In conclusion, rHuEPO and ACEIs can influence the total lymphocyte count or lymphocyte subset counts--the natural changes being disturbed with prolongation of CAPD treatment. The possibility of this influence should be taken into account when evaluating lymphocyte counts as indices of nutrition and immune status.

Adult↗