Characteristics of Epstein-Barr virus transformed B cell lines from patients with Alzheimer's disease and age-matched controls.

The characteristics of B cell lines isolated from patients with Alzheimer's disease (AD) and age-matched controls were investigated after having been transformed by Epstein-Barr virus (EBV). After isolation of mononuclear blood cells and in vivo or in vitro EBV infection, 35 and 21 lymphoblastoid cell lines (LCLs) were generated from 19 patients with AD (mean age 79.4 years) and 21 age-matched controls (mean age 80.0 years), respectively. B lymphocytes from AD patients were immortalised more easily than those from controls; the percentage of in vitro EBV infected LCLs (B95-LCLs) obtained in the AD group was significantly higher (76.2% versus 33.3% in the control group) and the mean time required for establishment was significantly lower (20.2 and 21.9 days versus 26.7 and 60.9 days in the control group). The EBV receptor and surface immunoglobulin (Ig) analyses showed no difference between the two groups. The expression of Epstein-Barr early antigens (EA) and viral capsid antigens (VCAs) revealed a tendency to higher viral replication in LCLs from AD patients; however, VCA expression remained limited to a small number of cells and did not affect overall cell growth. Finally, qualitative and quantitative differences were observed in the pattern of Ig production. Whereas spontaneously established LCLs from AD patients were generally monoclonal (80% of LCLs versus 33% in the control group), B95-LCLs were all polyclonal and secreted more IgM and IgA than those from controls; the mean IgM level was significantly higher in B95-LCLs from the AD group. These results suggest that B cells derived from AD patients seemed to be less differentiated than cells from age-matched controls.     

Radiosensitivity of lymphocytes from patients with Alzheimer's disease

The response after γ-irradiation of lymphocytes from 8 patients with Alzheimer's disease (AD), 3 patients with Huntington's disease and 13 normal subjects to stimulation by phytohaemagglutinin (PHA) was assayed by incorporation of [³H]thymlidine. The response of non-irradiated cells was found to be significantly lower in AD cells than in age-matched normals but not significantly lower in old normals than in young normals. However, the response of irradiated cells to PHA, expressed as a percentage of that in non-irradiated cells, was found to be similar in AD patients, young and old normals and in HD patients.     

Autologous Mixed Lymphocyte Reactions in Patients with Rheumatoid Arthritis and Juvenile Rheumatoid Arthritis: Both Non-T Cells and in-Vivo-Activated T Cells Can Act as Stimulator Cells

The lymphocyte responses in autologous mixed lymphocyte reactions (AMLR) between irradiated non-T and T lymphocytes from the peripheral blood (PB) of rheumatoid arthritis (RA) and juvenile RA (JRA) patients were decreased compared with the AMLR responses of normal PB lymphocytes. Normal AMLR responses were seen in the synovial tissue and the synovial fluid lymphocytes from RA and JRA patients. The lymphocyte responses were also decreased in AMLR between irradiated non-T cells from peripheral blood and T cells from synovial tissue (ST) in RA patients and between irradiated non-T from PB and synovial fluid (SF) T cells in JRA patients. However, when irradiated non-T cells from ST of RA patients or from SF of JRA patients were mixed with autologous PB T lymphocytes, increased lymphocyte responses were observed. SF T lymphocyters and ST T cells were also shown to stimulate autologous PB T lymphocytes.     

Fluorescent bodies in lymphocytes in chronic lymphocytic leukemia

The relative frequency of lymphocytes with nuclear Y-body-like fluorescent structures (F-bodies) was examined in ten patients with chronic lymphocytic leukemia (CLL) and in ten normal individuals. In each patient, the frequency was significantly higher as compared to that of normal controls. Female CLL patients showed a significantly higher proportion of F-body-positive (F-body +) cells than male CLL patients, whereas in normal males and females similar frequencies of F-body + cells were observed. There was no apparent relationship between the frequency of F-body + lymphocytes and the relative age of an individual. The presence of F-bodies in CLL had no obvious correlation with the presence of karyotypic abnormalities in these patients. Isolated T and B lymphocytes derived from four normal subjects showed no extra F-bodies as found in CLL patients.     

Blood lymphocyte subpopulations in breast cancer patients following radiotherapy.

Both T and non-T lymphocytes decreased immediately following radiotherapy in breast cancer patients. The relative depletion of non-T lymphocytes, however, was more marked than that of T cells. 3 years later the number and the proportion of non-T lymphocytes was higher than immediately after radiotherapy, while T lymphocytes were still depressed. The proportion of cells with membrane-associated Ig was higher in patients 3 years following radiotherapy than in non-treated patients and healthy controls. There was no difference in the proportion of T and non-T lymphocytes between patients with and without metastases, respectively.     

Autologous rosette formation in systemic lupus erythematosus

The number of autologous rosette forming cells was studied in patients with SLE. The effect of steroid hormones and of sera derived from SLE patients and normal controls on autologous rosette formation by lymphocytes of SLE patients and of normal controls was examined. The number of T lymphocytes capable of recognizing autologous red cells were found to be significantly lower in the SLE patients than in the group of normal controls. Steroid hormones, whether applied in vivo or in vitro, displayed an inhibitory effect on autologous rosette formation in SLE patients and normal controls alike. The effect in vitro was dose-related, and there was no difference in sensitivity between normal and SLE lymphocytes. Pooled sera of high immunocomplex content obtained from SLE patients were also found to reduce the number of auto-rosettes. Since the lymphocytes capable of recognizing autologous red cells are known to be post-thymic precursor cells equally subject to differentiation in helper and suppression direction and play an important role in the regulatory mechanism, reduction in their number or disturbances in their function and differentiation may well provide one of the factors accounting for the impairment of immune regulation in SLE.     

Monocyte function in Hodgkin's disease.

Monocyte functions were studied in 16 patients with Hodgkin's disease, 11 untreated and five in unmaintained complete remission. Eleven untreated patients with non-Hodgkin's lymphomas and 21 healthy persons were used as controls. Monocytes were isolated from peripheral blood and enriched to greater than 90%. Lymphoma monocytes showed normal ability to lyse human RBC coated with anti-D IgG antibodies as evaluated by a 51Cr-release assay. The ability of monocytes to augment or suppress concanavalin A stimulation of lymphocytes purified to greater than 98% was studied by incubation of a number of lymphocytes with increasing amounts of purified monocytes. The incorporation of 14C-thymidine was potentiated by a factor of 10 in the presence of equal amounts of monocytes. There was no difference between monocytes from Hodgkin, non-Hodgkin or healthy controls to augment patients' autologous or normal lymphocytes. Patient monocytes also suppressed the response at the same monocyte-lymphocyte ratio as normal monocytes. Stimulation of patient lymphocytes without the addition of monocytes was usually lower than that of normal control lymphocytes. The difference between patient and control lymphocyte stimulation was preserved in the presence of monocytes. It is concluded that monocytes from patients with active Hodgkin's disease or non-Hodgkin's lymphoma have normal helper and suppressor effects on patient or normal lymphocytes stimulated by Con A and normal antibody-dependent cytotoxicity.     

CD23/Fc epsilon R11 expression in contact sensitivity reactions: a comparison between aeroallergen patch test reactions in atopic dermatitis and the nickel patch test reaction in non-atopic individuals.

The immunopathology of patch test reactions to aeroallergens in patients with atopic dermatitis (AD) has been compared to that of contact sensitivity reactions to nickel in non-atopic individuals. Both reactions were found to exhibit equivalent erythema and induration on gross examination at 48 h. Four millimetre punch biopsies were obtained at 48 h frozen, and cryostat sections stained with a panel of MoAbs. The distribution of macrophages, dermal dendritic cells, Langerhans cells, T lymphocytes and the expression of CD23 antigen was recorded. Increased numbers of dermal dendritic cells, macrophages, T lymphocytes and Langerhans cells were found in the dermal infiltrates of both the nickel patch test reactions and the aeroallergen patch test reactions compared with their respective controls. There were no significant differences between atopic patch test reaction and nickel patch test reaction samples in the tissue distribution of these cell types. There was a significant increase in CD23 expression on Langerhans cells and dermal dendritic cells in the atopic patch test reactions, whereas an increase was only observed on dendritic cells in nickel patch test reactions. No significant difference in CD23 expression was observed in the control skin samples taken from patients with AD, nickel-sensitive patients and normal controls. This study supports the hypothesis that the aeroallergen patch test reaction in atopic dermatitis is a delayed hypersensitivity reaction, yet is distinct from the contact sensitivity reaction to nickel in terms of raised expression of CD23.     

The effect of a calf thymus acid lysate on bone marrow cell growth in vitro

Granulocyte-macrophage colony forming units (CFU-GM) were studied in cultures of bone marrow from 16 apparently healthy normal controls, 9 patients with the myelodysplastic syndrome, 5 patients with myeloproliferative disease and 2 with myeloma. Supernatants from non-stimulated 72 hr cultures of nonadherent mononuclear blood cells ("lymphocytes") stimulated the forming of an average of 38.4 colonies per 100,000 cells from normal marrow. The addition of GIBCO's commercial conditioned medium or of a medium produced by lymphocytes stimulated with different concentrations (5, 10 and 20 mcg/ml) of an acid lysate of thymus (thymomoduline), increased growth to 65.2 - 55.4 colonies (p less than 0.001 to 0.05). Similarly, a significant increase (p less than 0.05) was found in the number of clusters and colonies formed in cultures of marrow from patients with the myelodysplastic syndrome. In contrast, no growth was found when the thymus acid lysate was added directly to the bone marrow cultures, suggesting that the lysate induces the production of colony stimulating activity by lymphocytes, but does not contain it. Similarly no significant increase was found as regards the initially high number of colonies from the five patients with myeloproliferative disease, or as regards the initially low number in the two myeloma patients.     

Effects of gastric cancer cells on lymphocyte proliferation

Lack of lymphocyte infiltration into gastric cancer tissue appears to be an ominous prognostic indicator. The effects of gastric cancer cells on PHA-induced lymphocyte proliferation were studied. Peripheral lymphocytes were co-cultured for 72 hours with either gastric cancer cells or normal mucosal cells. Pairs of cancerous and normal mucosal cells from stomachs of eight patients, were separately co-cultured with peripheral lymphocytes either from patients or from normal volunteers. The degree of PHA-induced lymphocyte proliferation was measured by 3H-thymidine incorporation. The lymphocyte proliferation was inhibited by the presence of either gastric cancerous or normal mucosal cells in a dose-related manner. The lymphocytes from the normals proliferated twice as much as did the lymphocytes from the patients. The isotope incorporation occurred in lymphocytes rather than in gastric cells since the later incorporated insignificant amounts of isotope. There was no difference between gastric cancerous or normal mucosal cells inhibiting the proliferation of either normal or patients' lymphocytes (p greater than 0.05). In conclusion, gastric cancerous cells (up to 10(6)/ml) have no enhanced inhibition on lymphocyte proliferation when compared with normal gastric mucosal cells.     

Defective in vitro immunoglobulin production in response to pokeweed mitogen in patients with Hodgkin's disease pretreatment and in remission

In vitro production of IgG and IgM from peripheral blood lymphocytes and B-cell enriched fractions was assessed in a group of Hodgkin's disease (HD) patients and normal controls using pokeweed mitogen (PWM) stimulation. Our studies demonstrated a significant (P less than 0.01) reduction in the absolute number of helper (OKT4 positive) T cells and a significant alteration in the helper/suppressor T-cell ratio (0.89 +/- 0.15) compared to normal (1.83 +/- 0.31). Results from PWM stimulation experiments demonstrated that HD patients produced significantly lower IgG (P less than 0.01) and IgM (P less than 0.01) levels than controls. Synthesis of IgM but not IgG induced by PWM was subnormal after addition to patient B-cell cultures of autologous irradiated T cells or allogeneic irradiated normal T lymphocytes. Irradiated T cells from HD patients were as effective as normal T cells in helping PWM induced IgG and IgM synthesis by normal B cells. Our results suggest that in HD impaired circulating B-cell function is partly due to T-suppressor cell activity and furthermore that B-cell subpopulations producing different immunoglobulin isotypes may either be defective or vary in their susceptibility to T-cell suppression.     

Subpopulations of human helper and suppressor T lymphocytes

Mitogen driven differentiation of normal human mononuclear cells is a a well-established model for the study of antibody synthesis in man. In certain rare individuals who are clinically normal, unfractionated mononuclear cells or a mixture of purified B plus T lymphocytes differentiate into immunoglobulin producing cells in response to purified protein derivative of tuberculin (PPD) but not in response to pokeweed mitogen (PWM). To evaluate this observation we have irradiated T cells from such individuals to eliminate naturally occurring suppressor T cell activity and then added the irradiated T cells back to autologous B cells before culture. The B cells then responded to PWM. The original PPD responses of cells from these individuals were now significantly reduced. Although, there was no difference between PWM nonresponders and responders in the number of OKT-8 positive cells, elimination of OKT-8 positive cells in the PWM nonresponders with OKT-8 monoclonal antibody and complement resulted in a significantly increased response to PWM. This study indicates that there are suppressor T cells which specifically inhibit B cell response to PWM without affecting the PPD response. These results also show that the helper T cells involved in the PWM response are radioresistant and those involved in the PPD response are radiosensitive.     

Selective alterations in natural killer cell subsets in patients with atopic dermatitis.

We examined the immunophenotypic characteristics of natural killer (NK) cell subsets in patients with severe atopic dermatitis (AD), rhinitis allergica (RA) and in healthy controls. Expression of CD16, CD56 and CD57 antigens on peripheral blood lymphocytes was evaluated by simultaneous double immunocytofluorometry. Our results showed significantly lower percentages of cells with CD16, CD56 and CD57 surface antigens in patients with AD. Furthermore subdivision of the AD group into two subgroups, AD1 and AD0 (with and without antigen-specific IgE antibodies against potent inhalative allergens, i.e. mite, grass, rye, birch, cat) revealed that patients of subgroup AD1 showed a more prominent decrease compared to that of subgroup AD0. Moreover, we found a significant negative correlation between the percentage of CD56 + CD16 + NK cells and total IgE levels in serum, which were significantly higher in patients of subgroup AD1 than in AD0. NK cell activity was deficient in patients with AD but there was no difference between both subgroups. These data indicate that considerable heterogeneity in immunologic regulation may exist in patients with AD with regard to their NK cell subsets.     

Stimulating and differentiation factors for human B lymphocytes in systemic lupus erythematosus.

T cells from 18 untreated SLE patients produced significantly more B cell growth factor (BCGF) than did those from normal subjects. Those from SLE patients with active disease produced significantly more than did those from patients with inactive disease. The response to BCGF of SAC-stimulated B lymphocytes from SLE patients was higher than that of B lymphocytes from normal individuals. Similarly preactivated B cells from five of seven SLE patients also proliferated upon the addition of interleukin 1 (IL-1) whereas those of normal subjects did not. Simultaneous addition of IL-1 and BCGF had a synergistic proliferative effect on B cells from two of seven SLE patients but not on any of the controls. Interleukin 2 (IL-2) had no proliferative effect in either SLE or normal B cells. Supernatant fractions from T cells of seven of 10 patients with active SLE and three of 10 with inactive SLE induced more IgG production by CESS cells than did those of normal subjects indicating a higher production of B cell differentiation factor by SLE T cells than by those of controls. Our findings may explain the reported preactivation and predifferentiation of peripheral blood B cells from SLE patients and give insight into the mechanisms leading to the production of autoantibodies in this disease.     

Loss of suppressor T-lymphocyte function in patients with systemic lupus erythematosus (SLE).

Immunological reactivity in patients with SLE was studied in vitro trinitrobenzene sulphonate (TNP) specific antibody formation by peripheral blood lymphocytes. Lymphocytes from patients with SLE could produce an increased number of TNP-specific plaque-forming cells (PFC), while no such response could be seen in normal controls. Co-culture of lymphocytes from active SLE patients and normal controls was performed with TNP-Horse red blood cells (TNP-HRBC). The number of PFC by B lymphocytes from active SLE patients was suppressed by T lymphocytes from normal controls. On the other hand, the number of PFC by B lymphocytes from normal controls was increased by T lymphocytes from active SLE patients. Co-culture of lymphocytes from identical twins discordant for SLE was also performed, and the same results were obtained. We further examined the effects of Con A on antibody formation. Con A-treated T lymphocytes from a normal control markedly suppressed TNP-specific PFC by peripheral lymphocytes from active SLE patients. However, Con A-treated T lymphocytes from an active SLE patient did not suppress TNP-specific PFC by lymphocytes from another active SLE patient. These results suggest that active SLE patients showed a loss of suppressor T-lymphocyte function.     

Mitogenic responses to lipopolysaccharide by B lymphocytes from patients with systemic lupus erythematosus

Peripheral blood lymphocytes from 43 patients with systemic lupus erythematosus (SLE) and from age- and sex-matched normal controls were cultured with lipopolysaccharide (LPS) to examine the response to the polyclonal B-cell activator. Lymphocytes from active SLE patients incorporated 4840 +/- 471 (mean +/- SE) cpm in response to LPS, whereas lymphocytes from inactive SLE patients incorporated 6906 +/- 897 cpm. In contrast, lymphocytes from normal individuals incorporated 7452 +/- 1126 cpm. Ig synthesis of lymphocytes from active SLE in response to LPS stimulation was also less than that of normal individuals. The helper T-cell function of active SLE, as examined by co-culturing irradiated SLE lymphocytes with unirradiated normal lymphocytes, was normal. These results thus suggested that a defect of B lymphocytes exists in active SLE patients. This B-cell defect and T suppressor cells apparently play an important role in the pathogenesis of SLE.     

Mitogenic Responses to Lipopolysaccharide by B Lymphocytes from Patients with Systemic Lupus Erythematosus

Peripheral blood lymphocytes from 43 patients with systemic lupus erythcmatosus (SLE) and from age- and sex-matched normal controls were cultured with lipopolysaccharide (LPS) to examine the response to the polyclonal B-cell activator. Lymphocytes from active SLE patients incorporated 4840±471 (mean ± SE) cpm in response to LPS, whereas lymphocytes from inactive SLE patients incorporated 6906 ± 897 cpm. In contrast, lymphocytes from normal individuals incorporated 7452 ± 1126 cpm. Ig synthesis of lymphocytes from active SLE in response to LPS stimulation was also less than that of normal individuals. The helper T-cell function of active SLE, as examined by co-culturing irradiated SLE lymphocytes with unirradiated normal lymphocytes, was normal. These results thus suggested that a defect of B lymphocytes exists in active SLE patients. This B-cell defect and T suppressor cells apparently play an important role in the pathogenous of SLE.     

Silver-stained nucleolar organizer regions in bone marrow cells and peripheral blood lymphocytes of Philadelphia chromosome-positive chronic myelocytic leukemia patients

Silver-stained nucleolar organizer regions (Ag-NOR) in bone marrow cells and/or phytohemagglutinin-stimulated peripheral blood lymphocytes were compared between six normal healthy persons as controls and 22 Philadelphia chromosome (Ph)-positive chronic myelocytic leukemia (CML) patients, to examine if any disease associated changes occur in the expression of Ag-NOR. Although the frequency of Ag-NOR-positive cells and the number of Ag-NOR per cell were generally greater in lymphocytes than in bone marrow cells in both controls and CML patients, the Ag-stainability of these cell types in CML patients was considerably heterogeneous, compared with that found in controls. The peripheral lymphocytes of CML patients in the chronic phase, but not in the blastic phase, exhibited a significantly lowered Ag-stainability when compared with those of controls. while no such difference was observed between bone marrow cells of controls and leukemia patients in both phases of CML. In the blastic phase, however, the occurrence of Ag-NOR on the Ph of CML bone marrow cells was significantly less than expected. The present findings are discussed in relation to the existing data on the Ag-NOR expression in both normal and neoplastic cells.     

哮喘病人B细胞体外产生IgE过程中幼稚性T细胞的影响作用

目的：探讨支气管哮喘发病机制中Ｔ细胞的调节作用。方法：分离出病人辅助性Ｔ细胞中的幼稚性（Ｎａｉｖｅ）Ｔ细胞亚群即ＣＤ４＾＋ ＣＤ４５ＲＡ＾＋Ｔ细胞亚群,并将其与各自的Ｂ淋巴细胞共同培养。同时设定非特异性刺激原ＰＷＭ（美洲商陆）刺激组在刺激组,测定培养上清液中ＩｇＥ含量。结果：非刺激组病人ＩｇＥ含量明显高于健康对照（Ｐ〈０．０１）,刺激组病人与健康对照ＩｇＥ含量无显著差别（Ｐ〉０．０５）。结论：支气