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PubMed2004

Age-dependent variation in the proportion and number of intestinal lymphocyte subsets, especially natural killer T cells, double-positive CD4+ CD8+ cells and B220+ T cells, in mice.

The age-dependent variation in the proportion and number of lymphocyte subsets was examined at various extrathymic sites, including the liver, small intestine, colon and appendix in mice. In comparison with young mice (4 weeks of age), the number of total lymphocytes yielded by all tested organs was greater in adult (9 weeks) and old (40 weeks) mice. The major lymphocyte subset that expanded with age was interleukin-2 receptor (IL-2R) beta+ CD3int cells (50% of them expressed NK1.1) in the liver, whereas it was CD3+ IL-2Rbeta- NK1.1- cells at all intraepithelial sites in the intestine. Although NK1.1+ CD3+ cells were present at intraepithelial sites in the intestine, the proportion of this subset was rather low. The ratio of CD4 to CD8 tended to decrease among natural killer T (NKT) cells and T cells at all intraepithelial sites in the intestine with age. A unique population of double-positive CD4+ CD8+ cells in the small intestine increased in old mice. B220+ T cells were found mainly in the appendix and colon, and the proportion of these T cells decreased in old mice. Conventional NKT cells were very few in Jalpha281-/- and CD1d-/- mice in the liver, while NKT cells which existed in the appendix remained unchanged even in these mice. This was because unconventional CD8+ NKT cells were present in the intestine. The present results suggest that despite the fact that both the liver and intraepithelial sites in the intestine carry many extrathymic T cells, the distribution of lymphocyte subsets and their age-associated variation are site-specific.

Aging↗

PubMed2005

[Expression and clinical significance of inducible co-stimulator on peripheral blood T lymphocyte subsets in patients with systemic lupus erythematosus].

OBJECTIVE: To investigate the expression of inducible co-stimulator (ICOS) and other immunological molecules on peripheral blood T lymphocyte subsets in patients with systemic lupus erythematosus (SLE) and to find out the relationship with disease-activity, disease-stage and the contents of anti-dsDNA antibody and immunoglobulin in serum so as to pave the way for further studying the possibly immunologically pathological role of ICOS in SLE. METHODS: Peripheral blood samples were collected from 51 patients with SLE, 3 males and 22 females. Three-color flow cytometry was used to detect the levels of ICOS, CD45RO, CD45RA, and HLA-DR expression on the peripheral blood T lymphocytes subsets. The results were analyzed along with the disease-activity, disease-stage, contents of anti-dsDNA antibody and immunoglobulin in serum. Thirty healthy subjects were used as controls. RESULTS: Compared with the healthy subjects the level of ICOS expression on the peripheral blood CD4+ and CD8+ T cells in the patients with SLE during active- and stable-stages were significantly increased (all P < 0.05), but there was no significant difference between the last two group patients (P > or = 0.05); In the same patients, the level of ICOS expression on the peripheral blood CD4+, CD8+, CD45RO+, CD4+CD45RO+ and CD8+CD45RO+ cells in active-stage were significantly increased compared with those in stable-stage (P <0.05); The level of ICOS expression on the peripheral blood CD45RO+ cells in the untreated primary patients was higher than those with disease-relapse (P <0.05); The levels of ICOS expression on the peripheral blood CD45RO+ and CD4+CD45RO+ cells were significantly increased in the patients with serum anti-dsDNA antibody(+) and the patients with aberrantly high content of immunoglobulin compared with those of the patients with serum anti-dsDNA antibody(-) and the patients with normal content of immunoglobulin, respectively (all P < 0.05). CONCLUSION: ICOS is aberrantly highly expressed on certain peripheral blood T lymphocyte subsets in patients with SLE, which is related to disease-activity, disease-stage and the contents of serum anti-dsDNA antibody and immunoglobulin, thus ICOS may play a role in SLE pathogenesis.

Adult↗

PubMed1992

Unaltered lymphocyte subsets in hepatitis C virus-seropositive blood donors.

Lymphocyte subsets were evaluated by dual-color flow cytometry in whole blood specimens from 35 blood donors who were seropositive on enzyme-linked immunosorbent assay (ELISA) for hepatitis C virus (HCV) and whose sera reacted in a four-antigen recombinant immunoblot assay (RIBA) (referred to as the HCV+R group), 15 donors who were seropositive on ELISA for HCV with indeterminate or negative RIBA results (the HCV+I/N group), and 25 HCV-seronegative controls (HCV-group). The cell subsets assessed included natural killer cells, B cells, T cells, CD4 and CD8 subsets of T cells, and T-cell subsets defined by the coexpression of markers that appear (HLA-DR, CD25, CD38) or disappear (CD45RA) after activation. A one-way analysis of variance revealed no significant differences among the three study groups. These findings show that, unlike cytomegalovirus- and human immunodeficiency virus-positive individuals, HCV-positive individuals do not exhibit lymphocyte alterations indicative of the immune activation caused by chronic viral infection.

Antigens, CD↗

PubMed1984

Distribution of T-lymphocyte subsets in patients with vascular diseases.

The distribution of T-lymphocyte subsets was studied in patients with myocardial infarction, peripheral arterial disease and thrombophlebitis. The percentage of SRBC-rosette forming active - E[A] and total - E[T] lymphocytes in patients and healthy controls was not different. A significant increase in the percentage and number of theophylline-resistant - E[Thr] and T[M] - helper cells was found in patients with myocardial infarction or with thrombophlebitis, and a moderate elevation was observed in patients with peripheral arterial disease. No difference between patients and controls could be shown in the percentage and number of theophylline-sensitive E[Ths] and T[G] - suppressor cells. The imbalance of T-lymphocyte subsets is thought to be the consequence of sensitization against vascular tissues during vascular diseases.

Arterial Occlusive Diseases↗

PubMed2002

FACS monitoring of lymphocyte-subsets in patients with discoid and subacute-cutaneous lupus erythematosus receiving low-dose methotrexate.

OBJECTIVE: Our purpose was to describe and characterize the influence of low-dose methotrexate (MTX) treatment on absolute counts of peripheral blood cells and lymphocyte subsets in patients with cutaneous lupus erythematosus (LE). METHODS: In 6 patients with discoid (DLE) and 9 patients with subacute-cutaneous LE (SCLE) who received low-dose MTX (10-30 mg/week) leukocytes and lymphocyte subsets were monitored flowcytometrically. RESULTS: Absolute numbers of leukocytes, monocytes, lymphocytes, and lymphocyte subsets displayed cell-type specific patterns under low-dose MTX. Moreover, we found different reaction patterns in patients with DLE and SCLE. In patients with DLE leukocytes, lymphocytes, monocytes (p < 0.02), CD3+ cells (p < 0.04), CD3+ CD4+ cells (p < 0.009), and CD3+ CD8+ cells increase, and only B-cells decrease. Whereas in patients with SCLE no significant changes were found. CONCLUSION: Low-dose MTX treatment seems to independently influence peripheral blood cell compartments and thereby the underlying LE manifestation could be of some relevance.

Adult↗

PubMed1999

Interaction of tamoxifen with concurrent cytotoxic adjuvant treatment affects lymphocytes and lymphocyte subsets counts in breast cancer patients.

The effects and interaction of endocrine and cytotoxic adjuvant treatment on measures of cellular immunity were assessed in 41 stage I-II breast cancer patients from International Breast Cancer Study Group trials. Counts of lymphocytes and lymphocyte subsets [(T, T4, T8, B, natural killer (NK) and activated T (AT) cells] were assessed by flow cytometry immediately before adjuvant therapy at baseline and on day 1 of the 3rd cycle. Twenty-two patients received cyclophosphamide, methotrexate and 5-fluorouracil (CMF), 7 CMF and tamoxifen (TAM), and 12 TAM alone. On day 1 of the 3rd cycle the counts of total lymphocytes (P = 0.003) and all lymphocyte subsets (P<0.05) except AT cells were significantly lower than baseline in the CMF treatment group. There was no significant change in the CMF+TAM or in the TAM treatment group. The combination of CMF and TAM resulted in less pronounced decrease in lymphocyte and subset counts from baseline to day 1 of the 3rd cycle. It seems possible that there is an interaction between TAM with CMF that affects lymphocyte and lymphocyte subset counts during cytotoxic treatment.

Adult↗

PubMed1991

Reference ranges for lymphocyte subsets. A comparison of standard vs rapid whole-blood lysis techniques.

Reference ranges for lymphocyte subsets may vary with processing techniques, monoclonal reagents, or analytic methods. We compared reference ranges obtained for T- and B-lymphocyte subsets by means of standard manual whole-blood lysis with a wash step vs a rapid, no-wash whole-blood lysis system. Both techniques demonstrated reference ranges similar to those in previous literature reports. The ranges established with standard and rapid lysis were similar when antibodies directed to the same cluster designation were used. Although slight statistical differences in relative percentages of CD2 and CD3 lymphocytes were observed, these differences were probably not clinically significant. These data indicate that the rapid technique provides a standardized method for enumerating T and B lymphocytes in peripheral blood.

Adolescent↗

PubMed2006

[Effect of Chinese medicine for resolving dampness on activated and functional T lymphocyte subsets in chronic hepatitis B patients with dampness syndrome].

OBJECTIVE: To explore the relation between the effect of Huashi Decoction (HD) and levels of activated and functional T lymphocyte subsets in chronic hepatitis B (CHB) patients of dampness syndrome (DS). METHODS: Seventy CHB patients were nonrandomly assigned to two groups, the treated group (n = 45) treated by HD and the control group (n = 30) by nucleotide analogies. The clinical efficay was observed and levels of activated and functional T lymphocyte subsets were detected before and after treatment. RESULTS: The peripheral blood levels of CD8+ CD28+ and CD4+ CD28+ T cells were significantly lower in CHB patients than those in the healthy subjects (P < 0.01), which were higher in the treated group after treatment compared with those before treatment (P < 0.05), while the CD8+ CD38+ T cell level was significantly higher in CHB patients than in the healthy subjects (P < 0.05), which was lower in the treated and the control groups after treatment than those before treatment respectively (P < 0.05). No significant difference was found in the markedly effective rate and total effective rate between the two groups (P > 0.05). The levels of CD8+ CD28+ and CD4+ CD28+ T cells increased and the CD8+ CD3+ and CD4+ CD25+ T cell level decreased obviously after treatment in those who acquired marked effectiveness (P < 0.05), while they changed insignificantly in those who did not acquire marked effectiveness (P > 0.05). CONCLUSION: HD is effective in regulating activated and functional T lymphocyte subsets in CHB patients with DS, which might be its mechanism of inhibiting virus replication and eliminating virus.

Adolescent↗

PubMed1984

Lymphocyte subsets and immune complexes in long-standing diabetic patients: relation with the presence of microangiopathy.

Subsets of peripheral T lymphocytes by monoclonal antibodies and circulating immune complexes by two different methods were evaluated in 36 long-standing diabetic patients, 19 Type 1 (insulin dependent) and 17 Type 2 (non-insulin dependent). In all patients the presence of microangiopathy was assessed by retinal fluoroangiography, albuminuria and creatinine clearance. In patients with Type 1 diabetes a significant decrease of total T and of T cells with helper phenotype (T4), together with an increase of T cells with suppressor/cytotoxic phenotype (T8), were observed. No significant modifications in the percentage of T lymphocyte subsets were detected in patients with Type 2 diabetes. Immune complexes were found to be significantly increased in Type 1 compared with Type 2 diabetic patients. Patients with very high levels of T8+ cells did not have detectable immune complexes and had no evidence of microangiopathy. By contrast, patients with normal levels of these cells were found to have raised immune complexes and showed retinopathy of varying degree. The results of this study indicate that: (1) a relationship exists between cells with T8 phenotype, some immune complexes and the presence of microangiopathy; (2) the decrease of T4+ cells in Type 1 diabetics with long duration of disease may be responsible for the known susceptibility to infections in these patients.

Adolescent↗

PubMed1997

Circadian rhythm of leucocytes and lymphocytes subsets and its possible correlation with the function of the autonomic nervous system.

There are physiological variations in the levels of leucocytes. Among these, the circadian rhythm is very important in terms of the magnitude. Since newly identified lymphocyte subsets (i.e. extrathymic T cells) have recently been detected, a comprehensive study of the circadian rhythm was conducted. All leucocytes were found to vary in number or proportion with a circadian rhythm and were classified into two groups. One group--granulocytes, macrophages, natural killer (NK) cells, extrathymic T cells, gammadelta T cells, and CD8+ subset--showed an increase in the daytime (i.e. daytime rhythm). The other group--T cells, B cells, alphabeta T cells, and CD4+ subset--showed an increase at night. Humans are active and show sympathetic nerve dominance in the daytime. Interestingly, granulocytes and lymphocyte subsets with the daytime rhythm were found to carry a high density of adrenergic receptors. On the other hand, lymphocyte subsets with the night rhythm carried a high proportion of cholinergic receptors. Reflecting this situation, exercise prominently increased the number of cells with the daytime rhythm. These results suggest that the levels of leucocytes may be under the regulation of the autonomic nervous system.

Adult↗

PubMed1989

Lymphocyte subset analysis to predict progression to AIDS in a cohort of homosexual men in San Francisco.

A group of 10 leukocyte and lymphocyte subsets were measured by simultaneous dual immunofluorescence and flow cytometry in a group of homosexual men from the San Francisco General cohort. Absolute numbers and percentages of lymphocytes were determined in 30 individuals who progressed to AIDS and 29 who did not over a 44-month period at annual intervals. At entry into the study, all subjects were asymptomatic, HIV seropositive, and had multiple changes in lymphocyte subsets compared to HIV-negative controls. In progressors, large changes occurred from the first visit to the last visit before progression in both absolute numbers and percentages of CD4 cells. The percentage of HLA-DR-bearing CD8 cells also increased. We utilized a proportional hazards model to assign a predictive value for progression to AIDS to lymphocyte subsets in both univariate and multivariate tests. Increased DR-positive CD8 cells, decreased CD4 cells, and increased CD8-positive, Leu 7-positive cells independently predicted progression to AIDS at P less than 0.006 (relative hazard 5.8-4.0). In a multivariate model, the most useful tests were either increased numbers or percentages of DR-positive CD8 cells. These data suggest parsimonious approaches to following HIV-positive individuals and further support the possibility of autoreactive T cells in the pathogenesis of HIV-associated diseases.

Acquired Immunodeficiency Syndrome↗

PubMed1994

[Two-color flow cytometric analysis of lymphocytic subsets in renal cell carcinoma].

The subsets of tumor infiltrating lymphocytes (TILs) in 17 patients with renal cell carcinoma were studied by two-color flow cytometry and immunohistochemistry. In grade 1 tumors, CD4-positive cells were significantly increased compared to grade 2 and 3 tumors (35 +/- 7% vs 25 +/- 10%) (p < 0.05). In 10 of 14 patients, CD8-positive cells were predominant over CD4-positive cells, resulting in less than one in CD4/CD8-ratio. Regarding CD4 cells in all 17 cases, the helper T-cells (CD4-positive and CD45RA-negative) were statistically predominant over the inducer T-cells (CD4-positive and CD45RA-positive) (p < 0.01). When CD8 cells were investigated in 16 out of the 17 cases, the cytotoxic T-cells (CD8-positive and CD11b-negative) were found also significantly predominant over the suppressor T-cells (CD8-positive and CD11b-positive) (p < 0.01). Immunohistochemically, the infiltrating pattern of TILs in the 17 patients was classified into cluster (N = 4), separate (N = 6), and mixed types (N = 7). The tumor in the cluster and mixed type had a tendency to be dominant in low grade tumors, and had the prominent CD8-positive cells (more than 90% were cytotoxic T-cells) over CD4-positive cells. However, no statistical significance was noted in this finding. The advanced tumor tended to show the single type of TIL infiltration. These results suggest that cytotoxic T-cells consisted of most of TILs in cluster and mixed types play an immunoreactive role against renal cell carcinoma.

Adult↗

PubMed1994

Reduction of escapee formation in flow cytometric analysis of lymphocyte subsets.

Most experienced flow cytometrists performing immunophenotyping of lymphocyte subsets are aware of the escape phenomenon, in which some positive cells are found outside a lymphocyte gate based on forward and right angle scatter. However, little information is available on the levels of escapees formed with different antibodies, the roles of fluorochromes and lysing agents, the mechanism explaining the phenomenon, or methods to reduce it. We thus performed a systematic analysis of the escapee phenomenon to clarify these issues. A panel of monoclonal antibodies, including a phycoerythrin (PE) conjugate and a fluorescein isothiocyanate (FITC) conjugate of the same antibody from one manufacturer, was used to treat whole blood specimens, after which red cells were lysed using 0.15 M ammonium chloride (AmChl). The percentages of gated lymphocytes expressing CD3, CD8, CD19, and HLA-DR, but not CD2, CD4, CD16, and CD25, were significantly lower in FITC-stained versus PE-stained preparations. Correlated analysis of green fluorescence and forward scatter showed that, on average, 18% of CD3+ events, 24% of CD8+ events, and 25% of CD19+ events were escapees when using the FITC conjugate. In dual color analysis, CD3+ escapees were positive for CD62-P, CD13, and CD14, indicating that the escapee events consisted of FITC-anti-CD3-coated lymphocytes complexed with platelet-coated myeloid cells. In studies of the role of lysing agent, essentially no escapees were found in specimens treated with FACS lysing solution, which contains formaldehyde. We therefore included a similar denaturing agent, paraformaldehyde (0.1%), in the AmChl lysing agent, and found that the occurrence of escapees was markedly reduced. These findings show that the escapee phenomenon occurs when using some FITC-conjugated monoclonal antibodies in conjunction with AmChl lysing agent, and can be reduced by inclusion of paraformaldehyde in the lysing agent.

Ammonium Chloride↗

PubMed1991

Effects of interferon on intrahepatic human leukocyte antigens and lymphocyte subsets in patients with chronic hepatitis B and C.

We investigated the effects of interferon therapy on hepatocyte human leukocyte antigen class I and class II antigen expression and intrahepatic lymphocyte subsets in patients with chronic viral hepatitis B (n = 11) and C (n = 10). Interferon-alpha was administered intramuscularly in doses ranging from 3 to 18 million international units daily for 4 wk. Liver biopsy specimens were obtained just before and immediately after treatment, and the specimens were stained by the indirect immunoperoxidase method for evaluation of human leukocyte antigen expression and lymphocyte subsets. Before therapy, no significant difference was noted between hepatitis B and C in human leukocyte antigen class I antigen expression on hepatocytes or in the lymphocyte subsets in the intralobular and portal areas. After interferon-alpha treatment, hepatocyte expression of human leukocyte antigen class I antigens and serum beta 2-microglobulin levels were virtually unchanged in chronic viral hepatitis C patients, but both were increased in chronic viral hepatitis B patients. Human leukocyte antigen class II antigens were not expressed during treatment. The mean number of intralobular CD3+ and CD8+ cells and the mean serum ALT level decreased significantly in chronic viral hepatitis C patients (p less than 0.05) but not in chronic viral hepatitis B patients. The mean number of intralobular CD4+ cells was unaffected by interferon therapy in both groups. In all 21 patients, the changes in CD8+ cell numbers paralleled the changes in serum ALT levels. Our findings suggest that T-cell cytotoxicity may play an important role in hepatocyte damage in both chronic viral hepatitis C and chronic viral hepatitis B and that the response to interferon-alpha differs in these two types of hepatitis.

2',5'-Oligoadenylate Synthetase↗

PubMed1992

Activation of CD4+ and CD8+ lymphocyte subsets by streptozotocin in murine popliteal lymph node (PLN) test.

Time-related and dose-related popliteal lymph node (PLN) enlargement and lymphocyte subset alterations owing to subcutaneous (s.c.) injection of streptozotocin (STZ) into the foot pad were determined in (C57BL/6 x DBA/2)F1 [B6D2F1] mice. Early cell activation and time-related changes in T and B lymphocyte subsets were monitored during the onset of STZ-induced lymphoproliferative reaction by flow cytometry and immunophenotyping of lymphocyte subsets stained with a panel of monoclonal antibodies. Examination of cell size and chromatin decondensing for T and B cell subsets revealed differences in their activation profiles during the early phase of STZ-induced lymph node enlargement. The kinetics of the reaction showed initial activation and proliferation of CD4+ cells associated with accumulation of CD8+ and B cells. Subsequently CD8+ cells were activated and proliferated, but there was no evidence of early or late B cell activation as shown by the lack of increase in cell size or nuclear decondensation and the low and transient synthesis of immunoglobulins. The activation characteristics for CD4+ and CD8+ cell subsets in STZ-induced node enlargement were found to be analogous with T cell activation in acute allogeneic graft-versus-host (GVH) reaction in which the F1 recipient differed from the parent at both class I and II MHC loci. Our data support a central role for T cell activation in the induction of STZ-related PLN enlargement and suggest that recirculatory host B cells can play a major role in early node enlargement.

Animals↗

PubMed2005

Lymphocyte subsets and mood states in panic disorder patients.

This study was conducted to examine lymphocyte subset counts and mood states in panic disorder patients. Twenty patients with panic disorder and 20 age- and gender-matched normal healthy subjects were recruited for the study. We used the Spielberger State (STAIS) & Trait (STAIT) Anxiety Inventory, Hamilton Depression Rating scale (HAMD) and Hamilton Anxiety Rating scale (HAMA) to measure mood states in all subjects. Lymphocyte subsets counts were made by flow cytometry. Panic patients showed significantly higher scores for anxiety and depression than normal subjects. Panic patients showed no differences in terms of the numbers of immune cells, as compared with normal healthy subjects, other than a lower proportion of T suppressor cells and a higher T helper cell/T suppressor cell ratio. HAMA and STAIS scores were common factors that could predict T cell numbers and proportions, T helper cell numbers, and natural killer cell proportions in panic disorder patients. We suggest that anxiety levels are related to the T-cell population in panic disorder patients and that quantitative immune differences may reflect altered immunity in this disorder.

Adult↗

PubMed2000

[Cytomegalovirus infection and the changes of T-lymphocyte subsets in RAU patients].

OBJECTIVE: To study the quantitative changes of the T-lymphocyte subsets in RAU patients with different stages and to know the influences of cytomegalovirus(CMV) on cellular immunity in RAU patients. METHODS: CD4(helper/inducer) and CD8(suppressor/cytotoxic) cells in the peripheral blood were tested with monoclonally immunocytochemical technique (APAAP method) in 26 ulcerative stage patients and 10 inactive patients. RESULTS: The percentages of CD4 and CD8 cell were 36.19% and 29.38% respectively in ulcerative stage. The ratio of CD4/CD8 was 1.28 and it was inverted (< 1.0) in 26.92% of the patients. The percentages of CD4, CD8 cells was 39.90% and 34.21% in inactive stage respectively. The ratio of CD4/CD8 was 1.17. The number of CD4 and CD8 cells in ulcerative stage and CD4 cells in inactive stage was significantly lower than the normal values, P < 0.05. The number of CD8 cells in inactive stage was not significantly different compared with the normal value. However the numbers of CD3(total T cells), CD4 and CD8 cells increased to some extent in inactive stage. CD4 cells was significantly lower in 11 human cytomegalovirus (HCMV) DNA positive patients than that in 9 HCMV DNA negative patients (P < 0.05). CONCLUSIONS: A permanent disorder of cellular immunity was an important part of pathogenesis in RAU. CMV infection may be an important factor resulting in the disorder of cellular immunity because of the imbalance of T lymphocyte subsets.

Adolescent↗

PubMed1992

Kinetics of lymphocyte subsets from peripheral blood during a Plasmodium falciparum malaria attack.

Variations of lymphocyte subsets were followed longitudinally in 16 patients during an acute falciparum malaria attack. Before treatment, lymphocyte numbers were highly reduced, but the subset distribution was similar to that of healthy individuals. After parasite clearance, lymphocyte counts were normalized and the subset distribution was unchanged. This led to a normalization of all the absolute counts of lymphocyte subsets, except CD8+. The fast normalization of lymphocyte counts suggests that the initial decrease in lymphocyte numbers may reflect sequestration. Magnitude and kinetics of this variation were not related to parasite density or to severity of the attack. Activated T cells (CD3+HLA-DR+) were fewer in African than in European patients, suggesting the importance of the past exposure to malaria parasites in reallocation phenomena. These variations in lymphocyte numbers must be taken into account in the design and the analysis of cellular investigations in patients experiencing a falciparum malaria attack.

Adult↗