Activation of swine peripheral blood lymphocytes with human recombinant interleukin-2.

These studies examined the effect of the lymphokine interleukin-2 (IL-2) on porcine peripheral blood lymphocytes (PBL). Cultures of porcine (Yorkshire) PBL in human recombinant (r) IL-2 had a time-dependent increase in activation of killer cells. Previous reports of porcine killer cells derived from PBL indicated maximal lysis 18 hr after killer and target cells were mixed, with very little lytic activity at 4 hr. After culturing for 2 days in rIL-2, the cells acquire enhanced lytic capabilities resulting in substantial target lysis in the 4-hr assay. Enhanced lytic activity in the 18 hr assay occurs as early as 30 min after exposure to rIL-2, with a more significant increase after 2 days' exposure. The IL-2-treated cells have an increase in the incorporation of tritiated thymidine and an increase in percentage of granular cells. Tumour targets, such as Daudi, which are resistant to lysis, and L929, Mdt4 and P815, which are moderately susceptible to lysis by untreated swine PBL, had a substantial increase in lysis in IL-2-treated swine PBL. These results indicate human rIL-2 induces functional and morphological changes in swine peripheral blood lymphocytes.     

火把花根片对重症肌无力患者外周血淋巴细胞的影响

目的：探讨火把花根片治疗重症肌无力的免疫调节机制。方法：44例重症肌无力患者随机分为治疗组和对照组，两组均用溴吡斯的明作基础治疗，治疗组加用火把花根片治疗。在治疗开始前及治疗后一月抽血查外周血T淋巴细胞亚群。结果：治疗前治疗组CD4^ 、CDl9^ 细胞百分率、CD4^ ／CD6^ 比值和CD6^ 、CDl6^ ，56^ 细胞百分率与对照组比较无显著差异；治疗后治疗组CD4^ 细胞百分率、CD4^ /CD8^ 比值和CDl9^ 细胞百分率下降，CD8^ 细胞百分率增高，与对照组相比均有显著差异，而CDl6^ ，56^ 细胞百分率无明显变化。结论：火把花根片可以通过调节重症肌无力患者外周血T淋巴细胞亚群分布的方法来治疗重症肌无力。     

重症肌无力患者血清miR-126、Foxp3 mRNA的表达及临床意义

为探讨重症肌无力(myasthenia gravis,MG)患者血清miR-126、叉头翼状螺旋转录因子3(forkhead box P3,Foxp3)mRNA的表达及临床意义,为MG发病机制研究提供更多的依据,选择63例MG患者为MG组,50例体检正常者为对照组。采用RT-PCR检测血清miR-126、Foxp3 mRNA的表达水平,并采用Spearman相关性分析法分析miR-126与Foxp3 mRNA的相关性。结果显示,MG组患者血清miR-126的相对表达量(2.59±0.84)高于对照组(1.05±0.73,P0.01),而Foxp3 mRNA的相对表达量(1.58±0.81)低于对照组(3.15±0.96,P0.01);miR-126核苷酸序列在人、小鼠、狗等7个物种间存在高度保守性;MG组患者血清miR-126与Foxp3 mRNA呈负相关(r=-0.852,P0.01)。提示MG患者存在miR-126及Foxp3 mRNA的差异表达,且高表达的miR-126可能通过抑制Foxp3 mRNA的表达参与MG的致病。     

Subpopulations of T lymphocytes in myasthenia gravis patients.

Subpopulations of human peripheral blood T lymphocytes were examined in twenty-three myasthenic patients. T lymphocytes bearing receptors for the Fc portion of IgG (T gamma) were significantly increased in a third of the patients examined. T lymphocytes bearing receptors for the Fc portion of IgM (Tmu) were within normal values in all but two patients. Possible implications of these cells in the pathogenesis of myasthenia gravis are discussed.     

Follow-up of soluble interleukin-2 receptor levels after thymectomy in patients with myasthenia gravis.

Soluble interleukin-2 receptor (sIL-2R) levels were followed up after thymectomy by a quantitative immunoradiometric assay in 59 patients with myasthenia gravis (MG). Increased levels of sIL-2R were found in 30.5% of the patients before thymectomy. Serum levels were significantly higher in severely affected patients. Sequential sampling after thymectomy indicated a significant and progressive decline of sIL-2R levels within 2 years after surgery, which was well associated with clinical improvement or remission. The sIL-2R purified from sera of patients with MG had a molecular mass of 45 kDa as the normal sIL-2R. The decline after thymectomy of sIL-2R titers suggests a possible role of the thymus in the occurrence of sIL-2R in the periphery. Soluble IL-2R levels may represent a marker of disease severity in MG, which might be useful in the follow-up of individual patients.     

Immunomodulation by a dual altered peptide ligand of autoreactive responses to the acetylcholine receptor of peripheral blood lymphocytes of patients with myasthenia gravis

Myasthenia gravis (MG) is a T cell-dependent, antibody-mediated autoimmune disease. A dual altered peptide ligand (APL) that is composed of the tandemly arranged two single amino acid analogs of two myasthenogenic peptides was demonstrated to downregulate in vitro and in vivo murine MG associated autoreactive responses. Furthermore, treatment with the dual APL ameliorated the clinical manifestations of an established experimental autoimmune MG in mice. This study was undertaken in order to investigate the ability of the dual APL to immunomodulate MG-associated responses of peripheral blood lymphocytes (PBL) of patients with MG to the native autoantigen acetylcholine receptor (AChR). PBL of 22 of 27 patients with MG tested responded by proliferation to torpedo AChR. The proliferative responses of PBL of 21 of 22 responders were significantly inhibited by the dual APL. The inhibition was specific because a control peptide did not inhibit these proliferative responses. The dual APL also downregulated the levels of the secreted pathogenic cytokine IFN-gamma in supernatants of stimulated PBL of 80% of the tested patients. The latter inhibitions correlated with an upregulated production of the immunosuppressive cytokine, tumor growth factor beta. Thus, the results of our study demonstrate that the dual APL is capable of downregulating in vitro autoreactive responses of patients with MG and suggest that this peptide is a potential candidate for a novel specific treatment of patients with MG.     

Effects of pulsing procedure of interleukin-12 in combination with interleukin-2 on the activation of peripheral blood lymphocytes derived from patients with hepatocellular carcinoma

In patients with hepatocellular carcinoma (HCC), natural killer (NK) cell activity decreases significantly, and the reduced activity may be associated with the progression of HCC. In this study we evaluated the effects of pulsing with interleukin (IL)-2 and/or IL-12 on the activation of freshly isolated peripheral blood lymphocytes (PBL) derived from patients with HCC. PBL obtained from 9 HCC patients, 4 liver cirrhosis patients, and 9 normal subjects were cultured in the presence of IL-2 and/or IL-12. After 24 h of incubation, the levels of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha presented in the supernatants were determined by enzyme-linked immunosorbent assay (ELISA). The IFN-gamma and TNF-alpha production of PBL pulsed by a combination of IL-2 and IL-12 was significantly higher than those of PBL stimulated by either IL-2 or IL-12 alone. The mRNA encoding perforin, granzyme B, as well as IFN-gamma and TNF-alpha, were markedly enhanced in PBL stimulated with a combination of IL-12 and IL-2. The pulsing procedure of IL-12 in combination with IL-2 resulted in the increase of IFN-gamma and TNF-alpha, and the expression of perforin and granzyme B mRNA in PBL obtained from HCC patients, as well as in those obtained from normal subjects. These results indicate that adoptive immunotherapy based on PBL pulsed with a combination of IL-2 and IL-12 may be a promising adjunctive strategy for HCC treatment.     

协同刺激分子B7-H3在重症肌无力患者外周血中的表达

目的:检测重症肌无力(MG)患者外周血中协同刺激分子B7-H3的表达,并探讨其临床意义。方法:收集35例MG患者和44例健康对照(HC)外周血标本,采用免疫荧光标记,流式细胞检测外周血单个核细胞模型B7-H3(mB7-H3)的表达,ELISA法检测MG和HC血浆中可溶性B7-H3(sB7-H3)的含量。结果:MG患者外周血T淋巴细胞、单核细胞和B淋巴细胞上mB7-H3低水平表达,与HC无差异;MG患者外周血sB7-H3浓度为(2.166±0.958)ng/mL,显著低于HC(3.379±0.768)ng/mL;全身型MG(GMG)sB7-H3的浓度为(1.664±0.699)ng/mL,低于眼肌型MG(OMG)(2.396±0.985)ng/mL;并发胸腺异常的MG sB7-H3浓度为(1.593±0.441)ng/mL,低于胸腺正常MG(2.364±1.014)ng/mL;MGsB7-H3含量与QMGS评分呈负相关(r=-0.4189,P=0.012);免疫细胞mB7-H3与血浆sB7-H3无相关性。结论:MG患者sB7-H3表达下调,与疾病严重程度相关;在不同病理类型MG患者体内存在差异性表达,提示该分子可能参与了MG的免疫病理进程。     

Effect of interleukin-2 and mitogen on in vitro immunoglobulin production by peripheral blood lymphocytes from patients with selective IgM deficiency

Immunoglobulin production by mitogen and recombinant interleukin-2-(rIL-2)-stimulated lymphocytes from IgM-deficient patients was studied. The findings were that subnormal serum IgM levels did not necessarily predict defective in vitro IgG or IgM production, lymphocytes from some IgM-deficient patients exhibited defective T cell function, and rIL-2 did not enhance defective in vitro immunoglobulin production.     

Anti-acetylcholine receptor antibodies decrease after thymectomy in patients with myasthenia gravis. Clinical correlations

Clinical course and changes in serum levels of antibodies to the acetylcholine receptor (a-AChR) were followed in 82 patients with myasthenia gravis during a period of 1-8 years after thymectomy. Decrease in a-AChR immediately after thymectomy was influenced by changes in a total IgG. Immunosuppressive medication affected serum a-AChR at all points of time. In a subgroup of 41 patients without thymoma who had no immunosuppressive drugs, there was a steady decrease in a-AChR concomitant with clinical improvement from 6 weeks after thymectomy.     

Antiglobulin consumption test with sera from patients with myasthenia gravis

The antiglobulin consumption test was performed with sera from patients with myasthenia gravis, lyophilized skeletal muscle tissue serving as antigen.

Incubation of the saline-treated tissue preparation with the antiglobulin serum resulted in a small titre reduction of antiglobulin, indicating in vivo binding of γ-globulins to the tissue (basal consumption).

Treatment of the tissue with normal human serum resulted in a greater consumption. This was mediated by γG-globulins, normally present in human sera and with the capacity of binding to skeletal muscle (normal consumption).

Sera from twenty-four patients with myasthenia gravis were tested in the antiglobulin consumption test. A positive reaction (pathological consumption) was observed in twelve (50%). Cross-reactions with cardiac muscle and thymus were demonstrated.

     

Interleukin-2 receptor expression in peripheral blood lymphocytes from systemic lupus erythematosus patients: relationship to clinical activity

Deficient interleukin-2 (IL-2) production and other T-cell dysfunctions have been demonstrated in active systemic lupus erythematosus (SLE). The generation of IL-2 receptors is known to be important to the growth and differentiation of T and B lymphocytes. This study investigated IL-2 receptor expression in peripheral blood lymphocytes (PBL) from patients with active and inactive SLE. PBL from 27 SLE patients, diagnosed by revised ARA criteria, were assayed for IL-2 receptor expression, IL-2 and immunoglobulin (Ig) production. PBL from SLE patients with active disease spontaneously expressed increased numbers of IL-2 receptors compared to those with inactive disease (P less than 0.01) and normal donors (P less than 0.01). There was no significant increase in IL-2 receptors expression in PBL from active SLE patients in response to mitogenic stimulation with PHA compared to inactive SLE patients and normal donors. There was negligible IL-2 production in response to mitogenic stimulation and increased spontaneous IgG production by PBL from active SLE patients compared to normal donors (P less than 0.001). Purified B cells isolated from active SLE patients showed significant spontaneous IL-2 receptor expression when compared to spontaneous IL-2 receptor expression by normal B cells (P = 0.005). Therefore, in addition to derangements in Ig and IL-2 production, the level of spontaneous expression of IL-2 receptors may represent a cellular indicator of disease activity, and hence, may be a useful parameter in monitoring disease activity in SLE patients. The significance of the increased IL-2 receptor expression on B cells of active SLE patients is unknown, but may represent a marker of polyclonal activation of these cells.     

Responses in vitro of peripheral blood lymphocytes from patients with myasthenia gravis to stimulation with human acetylcholine receptor alpha-chain peptides: analysis in relation to age, thymic abnormality, and ethnicity

Peripheral blood lymphocytes (PBLs) were isolated from 24 patients with myasthenia gravis of three ethnic groups (Caucasian, African American, and Hispanic) and ten healthy individuals. We determined the in vitro proliferative responses of the PBL samples to each of 18 overlapping synthetic peptides corresponding to the entire main extracellular domain (residues 1-210) of the alpha-subunit of human acetylcholine receptor. The profiles of the T-cell responses (expressed in stimulation index [SI]) to the peptides varied among the 24 patient samples. There was a significant difference in the overall patient responses relative to controls toward 17 of 18 peptides. T cells from the patients gave responses greater than control mean SI + 4 standard deviation (Z(SI) > 4) to 2 approximately 9 peptides/sample. Six peptides, alpha 23-38, alpha 34-49, alpha 78-93, alpha 122-138, alpha 146-162, and alpha 182-198, were recognized with Z > 4 level by 42% to 58% of the patients' PBLs. The grouped patient responses, divided according to age, thymic diagnosis, or ethnicity, were compared with controls and with each other. Significant differences were observed between early- and late-onset cases in recognition of residues alpha 34-49 (p = 0.015) and alpha 78-93 (p = 0.053), and in recognition of residues alpha 12-27, alpha 56-71, alpha 134-150, and alpha 146-162 (0.0072 < p < 0.064) when two ethnic groups were compared with each other.     

Precipitation of auto-antibody in serum from patients with myasthenia gravis

Suitable means have been developed for the preparation of extracts of human or monkey skeletal muscle, which give precipitation with certain sera from patients with myasthenia gravis. These extracts are prepared in three solvents, namely, water, saline or sucrose. The water and saline extracts are the best for precipitation in fluid systems, conveniently carried out in capillary tubes. While all three types can be used for precipitation in gel diffusion, either double diffusion or immunoelectrophoresis, the sucrose extract is more convenient. In all these tests, the systems are to be incubated at 4°, and the results are to be read at 24 hours. This time limit is particularly important for the fluid precipitation, in order to avoid non-specific precipitation. With these precautions, it was found that no precipitation occurred with a large number of normal sera. Furthermore, only two out of about 100 sera from patients with various other diseases gave any precipitation. The precipitation antigen could not be found in extracts from a large number of other tissues, including heart, thymus and smooth muscle. Gel diffusion analysis revealed two major antigens and a minor antigen. Different precipitating myasthenic sera differ in their content of the respective antibodies. The antigenic material has not yet been chemically characterized, but the data suggest that it is not myosin or actomyosin.     

Increased interleukin-2 production by human peripheral blood lymphocytes treated with low doses of dexamethasone

Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 116, N ^o 7, pp. 68–70, July, 1993.     

Tg cells in peripheral blood lymphocytes from patients with rheumatoid arthritis

Using combinations of methods for detecting Fc receptors and B lymphocytes, non-B, non-T lymphocytes and T lymphocytes we have shown T_G cells to be significantly increased in rheumatoid arthritis (RA) patients' peripheral blood lymphocytes (PBL). The possible significance of this finding to the disease process is discussed.     

Antibodies to acetylcholine receptors in myasthenia gravis. In vitro synthesis by peripheral blood lymphocytes before and after thymectomy.

Pokeweed mitogen (PWM)-driven in vitro synthesis of antibodies to the acetylcholine receptor (PSA) was studied in non-thymoma patients with myasthenia gravis. In a group of 46 patients, the occurrence of PSA was related to the presence of the thymus or, in operated patients, the absence of a clinical effect of thymectomy. Sixteen patients were followed before and soon after thymectomy. PSA disappeared in all patients, at least temporarily, between 6 weeks and 1 year afterwards, independent of the clinical course and eventual clinical effect of the operation. A recurrence was found only in one of the five patients who derived no benefit from the operation. These findings support the hypothesis that the therapeutic effect of thymectomy can be explained by removal of a source of autoreactive lymphocytes. There was no correlation between the changes in serum levels of a-AChR and clinical improvement, suggesting a minor role of circulating peripheral blood lymphocytes (PBL) and the thymus in the total production of a-AChR.     

胸腺扩大切除治疗重症肌无力围术期血清IL-17、IL-18、IL-27水平分析

目的探讨胸腺异常合并重症肌无力（MG）患者行胸腺扩大切除术后外周血血清IL—17、IL-18、IL-27等细胞因子的变化情况及其意义。方法选取16例未经免疫抑制治疗的MG患者行胸腔镜下胸腺扩大切除术,采用双抗体夹心ELISA法检测术前及术后第7El外周血血清IL-17、IL．18、IL．27水平,以15例健康献血者的血清IL-17、IL-18、IL-27水平作为正常对照。结果胸腺切除术后MG患者外周血IL-17、IL-18、IL-27水平普遍降低,外周血IL-17、IL-18、IL-27水平在胸腺瘤和胸腺增生两种病理类型中无明显差异。结论手术切除胸腺可降低MG患者外周血IL-17、IL-18、IL-27水平。