Lymphocyte subpopulations and reactivity to mitogens in patients with scleroderma.

T lymphocyte subpopulations were studied in 40 patients with scleroderma (PSS), 26 of whom were studied simultaneously for lymphoproliferative responses to phytohaemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM). PSS patients exhibited a reduction relative to 42 age- and sex-matched controls in the absolute number and percentage of early E rosettes, late E rosettes and E rosettes formed with aminoethylisothiouronium bromide (AET) treated sheep red blood cells. There was no difference between patients and controls in the proportions of B lymphocytes. PSS patients exhibited normal lymphocyte transformation responses to PHA and ConA and an augmented response to PWM. The mitogen responses did not correlate with the absolute number or percentage of lymphocytes or T and B lymphocyte subpopulations. No correlation was observed between any immunological variable studied and the extent of skin or organ involvement, disease duration or therapy.     

B- and T-lymphocytes in toxic diffuse goitre.

Operation specimens of thyroid tissue from patients with Graves' disease were digested, with collagenase, down to a cell suspension. The lymphocytes in the suspension were concentrated by density-gradient sedimentation and the proportions of B and T lymphocytes were determined. Six patients with Graves' disease were studied. The proportions of B and T lymphocytes in the thyroid glands were similar to the proportions of B and T lymphocytes in the venous blood from healthy subjects.     

Immunomodulation by methimazole therapy in Graves' disease: rapid changes in activation stage of circulating regulatory T cell subsets, B cells and NK cells

In patients with Graves' disease, initiation of thyrostatic therapy with methimazole causes a selective reduction of thyroid but not other autoantibody levels. The mechanism of this immunosuppressive effect is unknown. In the present study, methimazole (20 mg daily) induced very rapid changes in the surface antigen expression of several circulating lymphocyte subpopulations in six patients with Graves' disease. Within 1 to 3 days of therapy, the proportions of HLA-DR+ cells within the CD8+(Leu 2+) subset (activated 'suppressor/cytotoxic' T cells), CD11+(Leu 15+) cells out of CD8+ cells ('suppressor' T cells), and CD45+R (Leu 18+) cells out of CD4+(LEU 3+) cells ('suppressor/inducer' T cells) increased significantly from 4.7 +/- 3.9% to 9.5 +/- 6.0%, from 7.5 +/- 1.5% to 17 +/- 5.8% and from 49 +/- 17% to 73 +/- 19%, respectively. In parallel, the percentages of DR+ cells within the CD4+(Leu 3+) subset (activated 'helper' T cells), 4F2+ cells out of CD19+(Leu 12+) cells (activated B cells) and 4F2+ cells out of CD16+(Leu 11+) cells (activated 'natural killer'-like cells) declined significantly from 7.2 +/- 5.6% to 2.8 +/- 2.1%, from 7.2 +/- 1.5% to 4.0 +/- 2.8% and from 5.5 +/- 3.5% to 2.8 +/- 4.9% at 3 days, respectively. In contrast, no consistent phenotypic changes occurred in lymphocytes drawn from six healthy subjects during 7 days of methimazole medication. Direct in vitro effects of methimazole on lymphocyte markers were not observed when blood mononuclear cells from untreated patients were incubated with either the drug (10(-4) and 10(-6)M) or with autologous pre/post treatment serum for 1 to 4 days. Methimazole thus induces rapid alterations in the subclass and activation marker expression of circulating lymphocyte populations in Graves' disease. These alterations are compatible with a down-regulation of B cell activity. Indirect evidence suggests that the thyroid gland is the source of secondary signals for these changes to take place.     

Canine cyclic hematopoiesis: alterations in T lymphocyte subpopulations in peripheral blood, lymph nodes, and thymus of gray collie dogs

Cyclic hematopoiesis (CH), also called cyclic neutropenia, is an inherited disorder known to occur in both humans and gray collie dogs. Previous reports have provided ample evidence to suggest that lymphocyte activity and regulatory mechanisms may be abnormal in CH. The present study examined the lymphocyte populations of several lymphoid compartments of gray collie dogs. The percentage of B lymphocytes in the lymph nodes of CH dogs was significantly increased whereas that of null lymphocytes was decreased. The percentage of T lymphocytes did not differ between CH and normal dogs, however, the proportions of T lymphocyte subpopulations were significantly different. The levels of T lymphocytes expressing IgGFc receptors (T gamma) in the thymus, lymph nodes, and peripheral blood were significantly increased; whereas the levels of T lymphocytes expressing IgMFc receptors (T mu) were significantly decreased. The percentage and absolute numbers of T gamma and T mu lymphocytes cycled in CH dogs. The percentage and absolute numbers of neutrophils were greatest when that of T gamma lymphocytes was reduced. The cycles of monocytes and T gamma lymphocytes occurred in close association and a linear relationship between the levels of these cells was observed both in terms of percentage (r = 0.62; P less than 0.01) and absolute number (r = 0.67; P less than 0.05). The percentage of T gamma and T mu lymphocytes were inversely correlated (r = -0.68; P less than 0.01).     

甲状腺自身免疫病浸润单人核细胞及甲状腺上皮细胞...

Specimens of thyroid tissue from 37 cases of autoimmune thyroiditis (AT), 13 cases of thyrotoxicosis accompanied with thyroiditis (TTOT), and 23 cases of Graves' disease (GD) were analyzed by immunohistochemistry with monoclonal antibodies. The majority of infiltrating mononuclear cells were learnt to be T cells. T+4 cells were abundant in lymphoid follicles and many T+8 cells were noticed in those areas with advanced destruction. B lymphocytes were predominantly located at the germinal centers. Most importantly, DR-positive thyroid epithelial cells were significantly increased with intense lymphocytes infiltration and severe destruction of the thyroid architecture. The percentage of T cell subpopulations, B cells, macrophages were somewhat similar. Anyhow, the increase of total number of infiltrating cells and the extent of inflammatory injury were remarkable in GD, TTOT and AT. The results support the idea that some GD cases may further develop in to autoimmune thyroiditis later.     

T lymphocyte subpopulations and Ia-positive T cells in patients with immunodeficiency.

T lymphocyte subpopulations (T gamma and Tmu) were studied in a group of 36 adult patients with immunodeficiency. Proportions and numbers of Ia(+) T cells were also studied in comparison to 46 normal adult controls. Values for per cent and total numbers of T gamma and Tmu cells indicated no uniform abnormality. Mean normal percentage of Ia(+) T cells was 2.4% whereas 16 to 29 immunodeficient patients showed elevated proportions and absolute numbers of Ia(+) T cells. Striking fluctuation in proportions of Ia(+) T cells was noted in serial studies of five immunodeficient subjects in contrast to similar analyses of normal controls. A correlation (P less than 0.01) was recorded between absolute numbers of Ia(+) T cells in immune deficiency patients and numbers of T mu cells. Depletion of T gamma cells by EA rosetting in patients with late-onset primary acquired hypogammaglobulinaemia did not result in significant change in IgG or IgM synthesis with T gamma-depleted T cells were co-cultured with normal B cells. Depletion of Ia(+) T cells likewise did not significantly influence Ig synthesis in co-culture with normal or immune-deficient B cells. These studies emphasize the complexity of defects present among any large group of patients with immune deficiency.     

Changes in lymphocyte subpopulations and CD3+/DR+ expression in sepsis

Objective   To detect lymphocyte subpopulations and CD3+/DR + expression in sepsis.
Methods   In a prospective clinical study we evaluated subpopulations of lymphocytes and percentage of CD3⁺/HLA-DR⁺ lymphocytes using two-color flow cytometry in 40 patients with sepsis and compared them with 34 healthy adults.
Results   Septic patients, when compared with healthy controls, have significantly lower percentage and absolute numbers of total T lymphocytes and CD4 T lymphocytes ( P  < 0.01). Absolute numbers of CD8 T lymphocytes, NK cells, CD3+/DR + lymphocytes and CD4/CD8 ratio were also decreased ( P  < 0.01). The percentage of B lymphocytes was increased ( P  < 0.01).
Conclusion   Our results are in agreement with previous findings in patients with sepsis after major surgery or trauma. The decreases in the percentage and absolute numbers of circulating lymphocyte subsets in non-surgical sepsis could represent a general reaction to stress. Increased percentage of B lymphocytes is most probably related to the bacterial etiology of the disease.     

Intrathyroidal lymphocyte subsets, including unusual CD4+ CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells, in autoimmune thyroid disease.

Intrathyroidal lymphocyte subsets were analysed in 13 euthyroid patients with autoimmune thyroid disease by two-colour flow cytometry and compared with subsets in peripheral blood. In both Graves' and Hashimoto's diseases, proportions of intrathyroidal CD5- B cells were higher than in peripheral blood. The numbers of such cells were correlated with serum levels of anti-thyroid microsomal antibodies. Proportions of T cells bearing alpha beta chains of T cell receptors (TCR alpha beta+ T; T alpha beta) and CD16+CD57+ natural killer (NK) cells were lower in the thyroid, but proportions of CD3hiTCR alpha beta-TCR gamma delta+ (T gamma delta) cells were not different. Proportions of CD4+Leu-8- helper T cells and CD4+CD57+ germinal centre T cells were higher and proportions of CD4+Leu-8+ suppressor-inducer T cells and CD8+CD57+ or CD8+CD11b+ suppressor T cells were lower than in the blood in both diseases. Proportions of CD5+ B cells were high in Graves' disease, and proportions of CD8+CD11b- cytotoxic T cells were high in Hashimoto's disease. Unexpectedly, CD4+CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells were present in thyroid tissues of both diseases. These findings suggest that: (i) an imbalance in the numbers of regulatory T cells and of NK cells that had appeared in the thyroid resulted in the proliferation of CD5- B cells, which were related to thyroid autoantibody production; (ii) CD5+ B cells and cytotoxic T cells are important for the different pathological features in Graves' and Hashimoto's diseases, respectively; and (iii) intrathyroidal CD4+CD8+ cells and CD3loTCR alpha beta lo/-CD4-CD8- cells may be related to the pathogenesis of autoimmune thyroid disease.     

SERUM IMMUNOGLOBULINS AND LYMPHOCYTE SUBPOPULATIONS DERANGEMENT IN TURNER'S SYNDROME

Abnormalities of the proportions of peripheral blood lymphocyte subpopulations and of immunoglobulin serum levels were found in twenty patients affected by Turner's syndrome. A slight but significantly decreased percentage of circulating T and B cells, an increased percentage of null cells and a decreased in vitro, responsiveness of lymphocytes to phytohaemagglutinin, concanavalin A and pokeweed mitogen were found in Turner's syndrome patients. IgG serum level was found significantly decreased in comparison with age-matched fifty-seven normal males and fifty-seven normal females and IgM serum level was intermediate between female and male values; Turner's syndrome patients with monosomy had an IgM serum concentration very close to male values. The derangement of T and B lymphocyte subpopulations, probably related to the aneuploidy, does not seem to be a severe one but it could account for the immunoglobulin abnormalities and for the association of Turner's syndrome with immunological disorders such as autoimmune diseases. The role of X chromosome on IgM serum level is discussed.     

Differences in effect of isoproterenol stimulation on levels of cyclic AMP in human B and T lymphocytes.

The level of cyclic AMP (cAMP) in human lymphocytes in the presence or absence of isoproterenol stimulation was studied in various lymphocyte subpopulations containing different proportions of B and T lymphocytes. Lymphocytes from thymus and peripheral blood (which contain a majority of T cells) were more sensitive to isoproterenol action than lymphocytes from tonsils or adenoids (where B cells are predominant). Using purified B or T lymphocytes from peripheral blood, tonsils or adenoids, we observed an absence of B lymphocyte response to isoproterenol, whereas T lymphocytes, which had a lower basal c-AMP level than B cells, exhibited in all experiments a significant increase in cAMP content after isoproterenol stimulation. The intensity of the response varied in the different T lymphocyte subpopulations.     

Decrease of peripheral large granular lymphocytes in Graves' disease.

Peripheral large granular lymphocytes (LGL) were enumerated in normal subjects and patients with autoimmune thyroid diseases. In normal subjects, the percentage of LGL was significantly lower in women (mean +/- s.d., 17.0 +/- 3.6%; n = 35; P less than 0.05) than in men (19.5 +/- 5.3%; n = 20). In untreated patients with thyrotoxic Graves' disease (GD), both the percentage (11.5 +/- 2.7%; n = 12; P less than 0.001) and the absolute count (244 +/- 102/mm3; P less than 0.01) of LGL were significantly lower than those in normal women (17.0 +/- 3.6% and 334 +/- 122/mm3; n = 35). No significant differences from normal controls were observed in the percentages or absolute counts of LGL in patients with euthyroid GD under treatment or in euthyroid or hypothyroid patients with Hashimoto's disease (HD). The percentage of LGL was inversely correlated with the serum levels of T4 and T3 and the free T4 index, but not with the titre of anti-thyroid antibodies, the size of goitre or the degree of proptosis in the group of untreated patients with GD and HD. The decreased levels of LGL in thyrotoxic patients with GD may be related to the self-perpetuation of thyrotoxicosis in patients with this disease.     

Immunohistological phenotyping of thyroid infiltrating lymphocytes in Graves'' disease and Hashimoto''s thyroiditis.

Subsets of lymphocytes in the thyroid were immunophenotyped by their surface antigens in frozen tissues of Hashimoto's thyroiditis and Graves' disease. Using triple layer immunoperoxidase staining (IP), monoclonal antibodies (T3, Leu 3, T8, anti-Tac and Leu 7) were employed to detect markers of T cell subsets, activated T cells, and a natural killer associated antigen. B cells were identified by 2 step IP with anti-IgD antisera. Excluding those cells forming lymphoid follicles, the density of lymphocytes infiltrating between thyroid epithelial cells was much higher in Hashimoto's thyroiditis than in Graves' disease. However, relative proportions of subsets were similar in both diseases. Most of the infiltrating cells were T3 positive T cells (T3+), with more T8+ (suppressor/cytotoxic T) than Leu 3+ (inducer/helper T). Some Leu 7+ were occasionally seen, but surface IgD positive mature B cells (IgD+) were almost absent. In contrast, IgD+ cells were densely aggregated in primary lymphoid follicles and mantle zones of secondary follicles. In these regions, Leu 3+ cells were about twice as frequent as T8+ cells. Some Leu 7+ and scarce Tac+ cells were also found. The present study indicates a major involvement of immunoregulatory T cells in autoimmune thyroid disease, and also suggested intrathyroidal maturation of B cells.     

Age-related changes in T- and B-lymphocyte subpopulations in the peripheral blood

Quantitation of T and B lymphocytes in infants and children is an important test in the diagnosis of a suspected immuno-deficiency. Previous studies indicated that the absolute and relative numbers of lymphocyte subpopulations vary with age, but these data in the pediatric age group are incomplete and often contradictory. We reviewed the literature and investigated the relationship between age and lymphocyte subpopulations in healthy infants and children using common methods and recent methodologic improvements. We found that absolute numbers of T and B cells followed the same trend as the total lymphocyte count, which was elevated at birth, increased in the first six months, and then gradually decreased to adult levels at approximately 13 years of age. Compared with adult values, the percentage of B cells also was higher at birth and continued to increase for six months, followed by a gradual decrease to adult levels by late childhood or early adolescence. The percentage of T cells gradually increased to adult levels by the same age range.     

Separation and analysis of mononuclear cells infiltrating the thyroid of patients with Graves' disease

A simple method was established for separating lymphocytes infiltrating the thyroid from thyroid epithelial cells. Namely, suspensions of minced thyroid from patients with Graves' disease were layered on a Percoll two-step density gradient (p = 1.050 and 1.077 g/ml) and centrifuged (400g, 30 min, 4 degrees C). In this way 0.1-18 X 10(5) lymphocytes/g of thyroid tissue with a purity of 65-95% were obtained. Thyroid lymphocytes were analyzed quantitatively with monoclonal antibodies by laser flow cytometry and compared with peripheral lymphocytes. The proportion of OKT3+ cells was decreased with increase in OKIa+ cells. The percentage of OKIa+ cells was significantly correlated with that of Leu12+ cells. The percentages of OKT4+ cells and OKIa+ cells were higher when analyzed with an extended gate window, which was arranged for detection of activated, large-sized lymphocytes. The percentages of OKT8+ and Leu7+ cells were not significantly different from those in peripheral blood. From these results it was concluded that the proportion of B lymphocytes is increased and that of T lymphocytes is decreased, the proportion of activated B lymphocytes is increased, some helper/inducer T cells are activated in the thyroid gland in Graves' disease, and these activated lymphocytes may be important in local production of antithyroid autoantibodies.     

Changes in T and B lymphocyte subpopulations before, during and after chemotherapy for malignant melanoma

The behaviour of T and B lymphocyte subpopulations before, during and after chemotherapy with DTIC or CCNU was studied in 87 melanoma patients. The effect on immune status of DTIC administered as adjuvant therapy was also reported. The immunosuppressive effect was found to be higher in patients with normal pretreatment T-cell values than in patients with low pretreatment values, and it was higher in metastatic than in nonmetastatic patients. Where T and B lymphocyte subpopulations were assessed in 10 metastatic patients after each course of chemotherapy, a progressive reduction of T-lymphocytes was observed, indicating a cumulative effect of the drugs with advancing course. The reduction of T lymphocytes was mainly due to the decrease of OKT4+ cells. A normalization of T lymphocyte subpopulations was observed after the suspension of therapy. No significant decrease in the number of B cells was observed in the DTIC treated metastatic patients, whereas a significative reduction in the absolute number was found in those treated with CCNU.     

The immunoregulatory disturbance in autoimmune thyroid disease

There is now considerable evidence for a genetically induced antigen-specific defect in suppressor T lymphocytes as the basis for AITD, derived from several laboratories and via different types of experimental techniques. In addition, there is now evidence for additive effects on reducing generalized suppressor T lymphocyte numbers and/or function by environmental factors as well as hyperthyroidism itself, and these effects would be superimposed upon the organ-specific defect. Such effects on generalized suppressor T lymphocyte numbers may act as precipitating and self-perpetuating factors, in Graves' disease at least. Presentation of the antigen by the thyroid cell via HLA-DR expression on its membrane does occur as a result of interferon gamma production by T lymphocytes. This in turn appears to be secondary to the initial specific immune assault and is not a primary inductive step. However, it may be important as an amplifying intermediate factor but cannot perpetuate the process in the absence of the underlying immune disorder. There is, however, no evidence for an underlying antigenic abnormality or stimulus in human autoimmune thyroid disease, and the initiating event would appear to be due to perturbation of the generalized immune system superimposed on the organ-specific abnormality. Variations in the serological and clinical expression of AITD would appear to depend on the severity of the original organ-specific disturbance in suppressor T lymphocyte function, plus the added factor of environmental influences playing upon generalized suppressor T lymphocyte function and numbers. Remissions in Graves' disease brought about by antithyroid drugs may well be via their effect on modulating the thyroid cell activity; this will then reduce thyrocyte-immunocyte signalling, allowing remission to occur in those patients with a partial organ-specific defect in suppressor T lymphocytes.     

T and B lymphocytes in patients with steroid-dependent bronchial asthma during long-term treatment.

Since corticosteroids are known to be a potent factor redistributing the peripheral blood lymphocytes to other body compartments, we estimated the sub-populations of T and B lymphocytes in peripheral blood of steroid dependent asthma patients. In twenty steroid-treated asthma patients and eighteen healthy blood donors the percentage and absolute number of lymphocytes forming spontaneous rosettes with sheep red blood cells ("early" and "late" T lymphocyte marker) and mouse red blood cells (B lymphocyte marker) were determined. The asthma patients were treated with 40 or 60 mg of triamcinolone acetonide (Kenalog) i.m. for at least one year or more prior to the study. No statistically significant differences were observed in lymphocyte subpopulations between steroid-treated asthma patients and healthy blood donors.     

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.     

T and B lymphocyte subpopulations and activation/differentiation markers in patients with selective IgA deficiency

Selective deficiency of immunoglobulin A (IgAD) and common variable immunodeficiency (CVID) are genetically closely related diseases, both of unknown pathogenesis. A plethora of abnormalities in lymphocyte subpopulations and expression of activation markers were repeatedly documented in CVID patients, while almost no data are available about lymphocyte subpopulations in IgAD patients. We determined basic lymphocyte subpopulations and those subpopulations that were reported to be abnormal in CVID patients (CD25, human leucocyte antigen (HLA)-DR CD45RA, CD45RO, CD27, CD28 and CD29 on both CD4(+) and CD8(+) cells, CD57 and CD38 on CD8(+) cells, CD21, CD27, IgM, IgD on B lymphocytes) in 85 patients with IgAD, 47 patients with CVID and in 65 healthy controls. Statistical analysis was performed by the Mann-Whitney U-test; significant P-values were determined by means of Bonferoni's correction. Our results showed an increase in the relative number of CD8(+) cells and a decrease in the absolute number of CD4(+) cells compared to healthy people, but similar abnormalities in CVID patients were much more expressed. IgAD patients had significantly decreased expression of HLA-DR and increased expression of CD25 on CD4(+) lymphocytes, also CD29 expression was decreased on CD8(+) cells, while other activation/differentiation markers on T cells (including the expression of CD45RA and CD45RO antigens) were not changed. There were no statistically significant abnormalities in B lymphocyte developmental stages in IgAD patients compared to healthy controls. Our observation showed that the majority of T and B lymphocyte subpopulation abnormalities described previously in CVID are not present in IgAD patients.     

Peripheral blood leucocytes in thyrotoxicosis (Graves' disease) as studied by conventional light microscopy.

Peripheral blood leucocyte counts in 104 thyrotoxic (Graves' disease) patients were compared with those in 107 normal subjects of similar age and sex. The thyrotoxic patients had a significant leucopenia with an absolute and relative neutropenia. They also had a relative lymphocytosis but the absolute lymphocyte count was within normal limits. The relative and absolute monocyte counts were normal.