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PubMed1995

Abnormalities of lymphocyte subsets in canine systemic lupus erythematosus.

Canine systemic lupus erythematosus (SLE) is a disease clinically very similar to its human counterpart. But so far, no study has reported an accurate evaluation of the lymphocyte subsets in the canine disease. Here, we present a study in which lymphocyte subsets have been evaluated in the peripheral blood of 20 dogs suffering from spontaneous systemic lupus erythematosus (SLE) in active and inactive phases, before and during treatment with prednisone and levamisole. 22 healthy dogs have been used as a control population. We show that canine SLE in active phases is associated with a several lymphopenia (1050 +/- 520 10(6) cells/l versus 2130 +/- 1 020 10(6) cells/l in controls). A striking finding is the imbalance of the CD4 and CD8 subsets (respectively 56.7 +/- 10.7% and 10.9 +/- 3.8% of CD4+ and CD8+ lymphocytes versus 40.5 +/- 11.5% and 18 +/- 4.4% in controls) and a strong activation of T-cells in active phases (64.1 +/- 16.9% of 2B3+ cells versus 46.5 +/- 16.7%). Moreover, we observed a persistence of the T subset imbalance during spontaneous evolution. In contrast, the treatment induced in dogs showing a good response the correction of CD4/CD8 ratio and no clinical manifestations, whereas in low responders no such improvements were observed. Thus, this work suggests that the main immunological imbalance seen in SLE could be associated with defective suppressor cells and provides further evidence of similarity of human and dog SLE.

Animals↗
PubMed1987

Flow cytometric analysis of lymphocyte subset phenotypes comparing normal children and adults.

The use of monoclonal antibodies and flow cytometry to characterize lymphocyte subsets has become an essential part of immunological research. In this study, we determined the normal ranges and absolute numbers of cells for several lymphocyte subsets in the peripheral blood of children and adolescents (ages 4-16 years). These were compared to the same subsets found in adults. Several differences were observed among the T and NK cell subsets. The total number of T cells, especially the CD4+ subset, were higher in children while two CD8+ subpopulations (CD8+CD11+ and CD8+Leu 7+) were found to be lower. Children had approximately half the absolute numbers of natural killer (NK) cells (CD16+), including a lower percentage of the CD16+CD8+ and CD16+Leu 7+ subsets. Monocytes and B-cell values were not significantly different between the two groups.

Adolescent↗
PubMed1989

Proliferation of lymphocyte subsets in the adult rat: a comparison of different lymphoid organs.

Adult, male Lewis rats received a single injection of 5-bromo-2'-deoxyuridine (BrdUrd) i.v. to label proliferating cells in the S phase of the cell cycle. After 1 and 24 h the thymus, bone marrow, blood, spleen, peripheral, cervical and mesenteric lymph nodes as well as Peyer's patches were removed. In cell suspensions surface staining was performed for B, T, T helper (Th) and cytotoxic/suppressor (Tc/s) T lymphocytes by identifying kappa light chain, CD5+, CD4+ and CD8+ cells, respectively. On the same slide the DNA label BrdUrd was demonstrated by a monoclonal antibody. B, T, Th and Tc/s lymphocytes proliferate locally both in central lymphoid organs such as the thymus and the bone marrow, and in peripheral lymphoid organs such as the spleen, lymph nodes and Peyer's patches. Within an organ the amount of proliferation among the lymphocyte subsets is similar, differing not more than threefold. Although concerning only a small fraction of cells within the organ, an unexpected finding is the high percentage of BrdUrd-labeled cells among B lymphocytes in the thymus (3%) and among T lymphocytes in the bone marrow (3%). One day after injection of BrdUrd the thymus contains 25% BrdUrd+ T lymphocytes, while the other organs investigated do not show more than about 2% BrdUrd+ B and T lymphocytes. Many of the newly formed lymphocyte subsets leave their organ of birth within 24 h. Thus the amount of proliferation in the lymphocyte subsets investigated is very similar and the differences between central (thymus and bone marrow) and peripheral lymphoid organs are much smaller than expected.

Animals↗
PubMed1993

[Lymphocyte subsets and interleukin-2 receptor in breast cancer patients].

The lymphocyte subsets and IL-2R (p55/Tac) in breast cancer patients were analyzed using monoclonal antibody and flow cytometry until 6 post-operative month. Results were as follows: 1. Lymphocyte fraction of CD3, 4, 8 was maintained from pre-operative to post-operative 6 month. 2. IL-2R value was lower in recurrence group than in breast benign disease group and it was lower. In pre-operative IL-2R in Stage III group than in Stage I and, Stage II. It was lower in DNA aneuploidy group than in DNA diploidy group. 3. At the post-operative 6 month, IL-2R was higher in Stage III group, ly-factor positive group, v-factor positive group and DNA aneuploidy group than in recurrence group. When breast cancer recurred after resection, IL-2R value returned to initial value. In conclusion, IL-2R may be possibly one of the factors of recurrence of breast cancer.

Adult↗
PubMed1996

Changes in lymphocyte subsets with age in perinatal cats: late gestation through eight weeks.

The age-related changes in numbers and proportions of peripheral blood lymphocyte subsets (pan-T, CD4, CD8, Ig, and null) were evaluated by two-color flow cytometry in 16 feline fetuses (56-58 days gestational age) and 21 kittens from birth through 8 weeks of age. Populations of pan-T+, CD4+, and CD8+ cells increased in total numbers as a function of increases in total lymphocyte numbers while proportions of these subsets remained relatively static. In contrast, both the total number and proportion of Ig+ cells increased from birth to 4 weeks of age, after which there were essentially no changes. Null (pan-T-, Ig-) cells were highest during late gestation and declined steadily thereafter to become a minimal component of the peripheral lymphocyte subsets. Compared with normal adult values, CD4/CD8 ratios were high throughout the 8 week study period. These results illustrate that the neonatal cat blood lymphocyte profile undergoes maturational changes and emphasize the importance of evaluating age-matched controls in studies of conditions that may alter feline lymphocyte subsets.

Aging↗
PubMed2001

Development of apoptosis and changes in lymphocyte subsets in thymus, mesenteric lymph nodes and Peyer's patches of mice orally inoculated with T-2 toxin.

Development of apoptosis and changes in lymphocyte subsets were examined mainly by flow cytometer in thymus, mesenteric lymph nodes and Peyer's patches of mice up to 24 hours after oral inoculation with T-2 toxin (10 mg/kg). T-2 toxin attacked Peyer's patches first, then mesenteric lymph nodes, and finally thymus in relation to the course of enteric absorption of orally inoculated T-2 toxin. The degree of lymphocyte apoptosis was prominent in the thymus, moderate in the Peyer's patches, and somewhat mild in the mesenteric lymph nodes, suggesting the difference in lymphocyte population susceptible to T-2 toxin. As to the changes in lymphocyte subsets, CD4+ CD8+ T cells were most sensitive to T-2 toxin, and CD4+ CD8- T cells were more severely depressed than CD4- CD8+ T cells in the thymus. In the mesenteric lymph nodes, CD3+ cells was more clearly affected than CD19+ cells, and the numbers of CD4+ and CD8+ cells were similarly decreased. In the Peyer's patches, the numbers of CD3+, CD 19+, CD4+ and CD8+ cells were unexceptionally decreased. In addition, among IgM+, IgG+ and IgA+ B cells, the number of IA+ B cells which are more important in the mucosal immunity was most severely affected.

Administration, Oral↗
PubMed2000

F cells, fetal hemoglobin levels, lymphocyte subsets, and frequency of crises in sickle-cell disease in Kuwait.

In vitro, fetal hemoglobin (Hb F) inhibits the aggregation of Hb S, and this may be clinically significant. Alterations in B and T lymphocytes seen in sickle-cell disease may be important in the immune deficit in these patients. We examined 16 patients with repeated crises (6 females and 10 males, age 9.3 +/- 3.4 years) and 15 patients with infrequent crises (9 females and 6 males, age 7.1 +/- 3.8 years) to determine the relationship between F cells, Hb F levels, lymphocyte subsets, and the frequency of crises. The proportion of CD2 and CD3 lymphocytes was significantly lower (P=0.015 and <0.0001, respectively) in both groups of patients than controls. CD4 lymphocytes were significantly lower (P = 0.018) and CD19 significantly higher (P = 0.007) than controls. CD45RO levels in both groups of patients were comparable with matched controls but significantly lower (P = 0.002) than adult values. Hb F levels in patients with and without frequent crises were comparable (P = 0.067). However, F cells in patients with infrequent crises were significantly higher than in patients with frequent crises (P<0.01). Alteration in the lymphocyte subsets did not correlate with the frequency of crisis in these patients.

Adult↗
PubMed1989

Lymphocyte subsets in human encephalitic and paralytic rabies.

Lymphocyte subsets of 7 patients with encephalitic and paralytic rabies were determined by immunocytochemical techniques using mouse monoclonal antibodies. Almost all patients had diminished mononuclear cells of Leu 7 phenotype (natural killer cells). Cells of Leu 12 marker (B cells) were decreased in 3 paralytic rabies patients compared with those of 4 patients in the encephalitic group.

Adolescent↗
PubMed1997

Lymphocyte subset redistribution during acute laboratory stress in young adults: mediating effects of hemoconcentration.

Acute psychological stress is known to alter the distribution of circulating lymphocyte subsets and also to cause a reduction of plasma volume. Data were reanalyzed from 4 previously reported studies (E. A. Bachen et al., 1995; T. B. Herbert et al., 1994; A. L. Marsland, S. B. Manuck, T. V. Fazzari, C. J. Stewart, & B. S. Rabin, 1995; A. L. Marsland, S. B. Manuck, P. Wood, et al., 1995) to determine the extent to which changes in the concentration of lymphocyte subsets are attributable to such hemoconcentration. Meta-analytic procedures showed circulating concentrations of T-suppressor/cytotoxic (CD8) and natural killer (NK) cells to increase following acute laboratory challenge, whereas T-helper (CD4) and B- (CD19) cell populations did not change. Adjustments for concomitant hemoconcentration reduced the magnitude of stress-related increases in CD8 and NK cells significantly and revealed a decrease in CD4 and CD19 cell concentrations from baseline to stress measurements. These data provide evidence (a) that increases in circulating numbers of CD8 and NK cells following acute stress are partially attributable to hemoconcentration and (b) that CD4 and CD19 cell concentrations decrease during acute stress when hemoconcentration is taken into account.

Adult↗
PubMed1986

Cryopreservation of human lymphocytes for assessment of lymphocyte subsets and natural killer cytotoxicity.

Cryopreservation of lymphocytes has become increasingly important, especially when the cells are to be used in retrospective studies of selected and dwindling populations, such as A-bomb survivors. This report describes an efficient method for cryopreservation of human lymphocytes which does not significantly alter various immunological characteristics of these cells. The proportions of Leu-1+ cells (T cells), Leu-2a+ cells (suppressor-cytotoxic T cells), Leu-3a+ cells (helper-inducer T cells), HLA-DR+ cells, Mo2+ cells (monocytes), B1+ cells (B cells), and Leu-7+ cells (natural killer (NK) cells), as determined by monoclonal antibodies, were found to be stable following cryopreservation. NK cell activity against K-562 target cells showed a 40-60% decrease immediately after thawing, but recovered to approximate pre-freezing levels after preincubation for 18 h. Neither lymphocyte subsets nor cell viability significantly changed following preincubation after cryopreservation. However, the ratio of cells binding to K-562 cells increased after this preincubation and may account for the observed recovery of NK cell activity. NK cell activity remained relatively stable up to 14 months of storage which confirms that freezing damage depends on the freezing process rather than on the duration of cryopreservation.

Antibodies, Monoclonal↗
PubMed1989

Lymphocyte subsets in kala-azar.

We examined lymphocyte subsets in peripheral blood from children with visceral leishmaniasis both in the active stage of the disease and after recovery. At diagnosis, CD2+ and CD4+ cells showed a significant decrease while CD8+ cells were significantly increased when compared with controls; CD4/CD8 ratio was inverted. All of these alterations returned to normal values 3 months after recovery.

Anemia↗
PubMed1986

The effect of hemodialysis on lymphocyte subsets during dialysis.

To clarify the effect of hemodialysis on immune response of dialysis patients, peripheral lymphocyte subsets were measured using monoclonal antibodies, OKT3, OKT4, OKT8, OKIal and Leu7. The absolute numbers of OKT3+, OKT4+, OKT8+ and OKIal+ cells were significantly lower in dialysis patients than controls. During dialysis, the relative proportions of OKT3+ and OKT4+ cells increased, and the absolute numbers of OKIal+ and OKT8+ cells decreased, 30 minutes after initiation of dialysis. At the end of the dialysis, the absolute numbers of OKT3+ and OKT4+ cells increased. The absolute number of Leu7+ cells showed low values during dialysis. These changes of lymphocyte subsets during dialysis may have influence on the immune response of dialysis patients.

Adult↗
PubMed1992

Activation of CD4+ and CD8+ lymphocyte subsets by streptozotocin in the popliteal lymph node assay. II. Comparison with acute graft-vs-host reaction in H-2 incompatible F1 mouse hybrids.

Changes in lymphocyte subsets during an acute GVH reaction were compared to STZ-induced PLN response in mice. The GVH reaction was induced locally by sc injection of parental C57Bl/6 [B6] spleen cells into (C57Bl/6 x DBA/2) F1 footpad [B6D2F1]. Early cell activation and time-related changes in T- and B-lymphocyte subsets were monitored during the onset of the GVH reaction by flow cytometry and immunophenotyping. Examination of cell size and chromatin decondensing for T- and B-cell subsets showed differences in activation profile during the early phase of the GVH reaction. The present study provides direct evidence for early in vivo activation of both CD4+ and CD8+ T-cells. Our data confirm the central role of T-cell activation in the induction of a GVH reaction and suggest that recirculatory host B-cells can play an important role in early GVH node enlargement. Overall, our comparative analysis supports the concept of polyclonal T-cell activation for both STZ-related and GVH-induced lymphoproliferation. Chemicals-induced lymphoproliferation leading to autoimmune reactions is a challenging issue. A number of drugs and chemicals have been tested in the PLNA assay for lymphoproliferative potential. We previously reported the activation and proliferation of T-cell subsets following STZ injection into murine footpads. The STZ-induced PLN enlargement and proliferation characteristics of T- and B-cell subsets were postulated to be similar to those of an acute allogeneic GVHR. In the present study, a cytometric analysis of T- and B-cell subsets in PLNs was performed during an acute allogeneic GVHR, for comparison purposes. Such a reaction results in a massive node enlargement five to ten times that seen after stimulation with conventional antigens. Acute GVHR is believed to be a direct consequence of the high frequency of alloreactive donor T-cells inducing a massive proliferation of B-cells, almost exclusively of host origin, in GVHR nodes. It is now widely accepted that donor T-cells activated as the result of exposure to foreign MHC antigens in the recipient, secrete various cytokines which assist the host B-cells and bypass the normal B-T cell cooperation. Induction of an acute GVHR, as in the parental B6--->recipient B6D2F1 model, requires the injection of CD4+ and CD8+ donor T-cells into an F1 recipient that differs from the parent at both MHC class I and II loci.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Changes of lymphocyte subsets in peripheral blood before and after operation of patients with advanced ovarian carcinoma.

Comparison was made between lymphocyte subsets in peripheral blood from patients with benign ovarian tumor and those with advanced ovarian carcinoma. In addition, changes of lymphocyte subsets of patients with ovarian carcinoma before and after operation were also examined. The percentage and absolute number of CD3-/HLA-DR+ (B cells) in peripheral blood from patients with advanced ovarian carcinoma were significantly lower than values from patients with benign ovarian tumor, whereas both percentage and absolute number of CD3-/HLA-DR- (null cells) cells in patients with advanced ovarian carcinoma were significantly higher. Although there was no significant difference in natural killer (NK) cell subsets (CD57+ CD16- and CD57+ CD16+ cells) between patients with benign ovarian tumor and ovarian carcinoma, the percentage and absolute number of CD57-/CD16+ (highly differentiated NK cells) cells in patients with ovarian carcinoma were significantly higher than those in patients with benign ovarian tumor. Both the absolute number and percentage of CD3+/HLA-DR+ (activated T cells) cells in ovarian cancer patients with minimal residual tumors after operation were significantly increased, compared to the levels before operation, while the values in the patients with large residual tumors were significantly decreased. In addition, the percentage and absolute number of CD3-/HLA-DR- (null cells) cells in the patients with minimal residual tumors were significantly decreased after operation, while values in the patients with large residual tumors remained unchanged before and after operation. The patients with minimal residual tumors after operation were characterized by a significant increase in the percentage of CD57- CD16+ (highly differentiated NK cells) cells. On the other hand, in the patients with large residual tumors no change of the NK cell subsets was observed before and after operation.

Adult↗
PubMed1993

Expression of decay-accelerating factor and CD59 in lymphocyte subsets of healthy individuals and paroxysmal nocturnal hemoglobinuria patients.

The expression of phosphatidylinositol (PI)-anchored complement-regulatory membrane proteins on circulating blood cells has been well clarified; however, the PI proteins on lymphocyte subsets have not been fully analyzed yet. We examined the expression of decay-accelerating factor (DAF) and CD59 on the T lymphocytes (CD2+, CD3+, CD4+, and CD8+) and CD20+ B lymphocytes in ten healthy volunteers and 12 paroxysmal nocturnal hemoglobinuria (PNH) patients by cytofluorometry. In healthy controls, each subset of lymphocytes showed a small population of cells weakly positive and a large population of cells strongly positive for DAF and CD59, while erythrocytes showed a single population of cells positive for the PI proteins. The two-population expression of DAF was most distinctive in CD8+ T cells among the subsets. In PNH, each subset of lymphocytes showed a moderately higher population of cells weakly positive and a smaller population of cells strongly positive for the membrane proteins compared with those in the healthy controls. Moreover, in some PNH cases, a negative population for the proteins was found in all subsets. Thus the analysis of PI-anchored proteins on lymphocytes subsets (especially CD8+ T cells) was considered to be of diagnostic value in PNH patients who receive blood transfusion after hemolytic attack of affected erythrocytes. Furthermore, the two-population expression of PI proteins in normal lymphocytes suggests that membrane PI protein would be a new subset marker of lymphocytes.

Adult↗
PubMed1986

[The alternation of peripheral lymphocyte subsets in head and neck carcinoma].

Peripheral lymphocyte subsets of 115 patients with head and neck carcinomas were examined by the monoclonal antibody technique. In fresh tumor-bearing, the OKT 4 rate was decreased (P less than 0.01) and the OKT 8 rate was increased (P less than 0.05). Consequently the OKT4/OKT8 ratio was decreased (P less than 0.01) compared with normal healthy individuals. This result shows the abnormality of cellular immunity in carcinoma cases. The OKT4/OKT8 ratio reflected the clinical stages and courses sufficiently and correlated with other immunological parameters. The OKT4/OKT8 ratio is considered to be one of the parameters for elucidating the clinical conditions and immunities of carcinoma patients.

Adenocarcinoma↗
PubMed1984

Prospective study of lymphocyte subsets in subjects genetically susceptible to type 1 (insulin-dependent) diabetes.

A prospective study of lymphocyte subsets has been carried out for 18 months in 58 healthy first-degree relatives of Type 1 (insulin-dependent) probands. Subjects selected for presence or absence of islet cell antibodies included 10 with complement-fixing islet cell antibodies, 10 with conventional islet cell antibodies and 38 without islet cell antibodies. Immunoregulatory and effector lymphocytes subsets, and in particular activated T-cells, were investigated using a panel of monoclonal antibodies. The results showed no significant changes in total T, helper, suppressor/cytotoxic cell or K/NK cells. Activated T-cells were observed at least once in 22 subjects using the 4F2 monoclonal antibody and in 11 using the Tac antibody. Seven subjects had 4F2-positive cells on repeated occasions and one twice showed Tac-positive cells. Fluctuations and/or loss of islet cell antibodies were observed during follow-up. There was no correlation between presence of activated T-cells and either islet cell antibody status of HLA haplotype sharing with the diabetic proband. On the other hand, a significant correlation was observed between HLA-DR3 positivity of subjects and the occurrence of activated T-cells (both 4F2-positive and Tac-positive). We conclude that subjects with HLA-DR3 may be especially prone to T-cell activation. As none of the 'high risk' individuals developed diabetes in the course of follow-up, the relevance of these observations in the pathogenesis of Type 1 diabetes needs more prolonged investigation.

Antibodies↗
PubMed1998

Lymphocyte subsets in cord blood of preterm infants: effect of antenatal steroids.

The objective of this study was to evaluate prospectively the influence of gestational age (GA) and short-term antenatal steroids on total lymphocyte count and lymphocyte subsets in cord blood from preterm infants. Two-color flow cytometric analyses of lymphocyte subsets were performed on cord blood collected from 67 infants. These infants were grouped according to GA: group I (term, n = 19); group II (GA 33-37 weeks, n = 25); group III (GA <33 weeks, n = 23). The mean absolute lymphocyte counts (ALC) in groups I, II and III were 5.6 +/- 2.5 x 10(3)/ microl, 4.3 +/- 1.5 x 10(3)/ microl and 3. 5 +/- 1.8 x 10(3)/ microl respectively. The mean values for CD4+ lymphocytes in groups I, II and III were 2.7 +/- 0.8 x 10(3)/ microl, 2.0 +/- 0.8 x 10(3)/ microl and 1.6 +/- 0.9 x 10(3)/ microl respectively. Mean values for CD8+ lymphocytes were 0.9 +/- 0.3 x 10(3)/ microl, 0.6 +/- 0.3 x 10(3)/ microl and 0.5 +/- 0.3 x 10(3)/ microl respectively. With decreasing GA, there was a statistically significant decrease in ALC (p = 0.0035), CD4+ lymphocytes (p = 0. 0013) and CD8+ lymphocytes (p = 0.0064). We then evaluated the effect of antenatal steroids, now routinely administered to women with preterm onset of labor to facilitate fetal lung maturation, and found that after adjusting for GA, infants of women on antenatal steroids had significantly fewer ALC (p = 0.0001), CD4+ lymphocytes (p = 0.02) and CD25+ lymphocytes (p = 0.03). In this population of infants, the decreased number of lymphocytes seen at younger GAs is associated with antenatal steroid use.

Betamethasone↗