Clinical impact of cyclosporine cellular pharmacodynamics in minimal change nephrotic syndrome

BACKGROUND: Cellular pharmacodynamics of cyclosporine (INN, cyclosporin) is considered to be closely implicated in clinical efficacy of the drug in kidney transplantation and other immunologic disorders. We applied this strategy to patients with minimal change nephrotic syndrome to predict individual clinical efficacy of cyclosporine. METHODS: Drug sensitivity tests were carried out with peripheral blood mononuclear cells from 31 patients with minimal change nephrotic syndrome. The 50% lymphocyte-mitosis inhibition of cyclosporine on in vitro blastogenesis of peripheral blood mononuclear cells stimulated with concanavalin A were estimated, and interpatient variations of 50% lymphocyte-mitosis inhibition were evaluated. The relationship between cyclosporine-50% lymphocyte-mitosis inhibition and clinical outcomes indicated a decrease of urinary protein and the period required for complete remission under cyclosporine therapy was examined in 14 patients. We also evaluated the correlation between cyclosporine-50% lymphocyte-mitosis inhibition and interleukin-2 production and percentages of interleukin 2 receptor-positive peripheral blood mononuclear cells in vitro. RESULTS: Cyclosporine 50% lymphocyte-mitosis inhibition on peripheral blood mononuclear cell blastogenesis deviated largely between patients from 0.2 to 86.0 ng/mL. We found a statistically significant negative correlation between cyclosporine-50% lymphocyte-mitosis inhibition in vitro and decreasing rates of urinary protein at 1 week after onset of cyclosporine administration (r = -0.655, P < .02). When we arbitrarily divide the 14 patients who received cyclosporine therapy according to their median 50% lymphocyte-mitosis inhibition of cyclosporine into two groups, that is, a high-sensitivity group (50% lymphocyte-mitosis inhibition < 18.1 ng/mL, n = 6) and a low-sensitivity group (50% lymphocytemitosis inhibition > 18.1 ng/mL, n = 8), the period required for complete remission was significantly shorter in the high-sensitivity group (P < .03). The 50% lymphocyte-mitosis inhibition of cyclosporine on interleukin-2 production in culture medium was correlated with 50% lymphocyte-mitosis inhibition of the drug on peripheral blood mononuclear cell blastogenesis (r = 0.806, P < .02). Decreasing rates of interleukin-2R-positive cells by cyclosporine treatment in vitro were negatively correlated with peripheral blood mononuclear cells blastogenesis in the presence of the drug (r = -0.694, P < .02). CONCLUSIONS: Peripheral blood mononuclear cell response to cyclosporine in vitro is closely related to clinical efficacy of the drug in minimal change nephrotic syndrome. Peripheral blood mononuclear cell resistance to cyclosporine was correlated with ability of the cells to express interleukin 2 and interleukin 2R.     

Diagnosis of viral myocarditis by cardiac magnetic resonance and viral genome detection in peripheral blood

In patients with acute myocarditis, viral genome can be detected in plasma and peripheral leukocytes. Its relationship with active myocardial inflammation, however, is not well understood. Myocardial edema as a feature of inflammation and myocardial necrosis or fibrosis can be frequently observed in patients with acute myocarditis by cardiovascular magnetic resonance (CMR). We assessed the association of viral genome presence in peripheral blood samples with myocardial edema and irreversible injury. We examined consecutive patients with clinically suspected myocarditis after an episode of viral illness. State-of-the-art methods were used for detecting myocardial edema and irreversible injury using CMR and viral genome applying reverse transcribed, nested polymerase chain reaction in peripheral blood samples. The specificity of viral amplification products was confirmed by automatic DNA sequencing. Of a total of 55 patients (53.5 ± 15.6 years), 21 were positive for viral genome in peripheral leukocytes. Interestingly, 18 (86 %) of these patients also showed global myocardial edema, as compared to only 7/34 (21 %) without PCR evidence for viral genome. The overall agreement between CMR criteria for edema and viral PCR was 84 %. In contrast, there was no significant relationship of viral genome presence with myocardial necrosis or scars. In patients with clinically suspected myocarditis, myocardial edema but not irreversible myocardial injury is associated with the presence of viral genome in peripheral blood.     

Rejection monitoring by lymphocyte blastogenesis assay in canine lung allotransplantation

Seventy-seven adult mongrel dogs underwent left lung transplantation to study the immunological detection of posttransplant rejection response using the lymphocyte blastogenesis assay. Experimental animals were divied into 3 groups; control autotransplantation (4 dogs), control allotransplantation (32 dogs) and immunosuppressed allotransplantation (41 dogs). The lymphocyte blastogenesis assay included mixed lymphocyte reaction (MLR), lymphocyte reactivity to lectins (PHA, ConA and PWM) and spontaneous blastogenesis in peripheral lymphocyte culture methods. The latter two tests were performed in serial blood samples up to the 38th posttransplant day. In the control autotransplant group there were no particular changes in posttransplant PHA reactivities. In the animals with lung allograft, results of preoperative MLR between donor and recipient showed no particular relationships to posttransplant rejection response. The recipient, however, showed a decreased response to PHA and an increased spontaneous blastogenesis at the period undergoing rejection which were confirmed by blood samples taken within 2 days before sacrifice of animals whose transplant specimens showed apparent histologic findings of rejection. In the immunosuppressed allotransplantation group, those findings of PHA response and spontaneous blastogenesis had often preceded the rejection episode detected on chest x-ray films, whereas there were no particular changes in both parameters of dogs bearing enhanced allografts. The serial detection of spontaneous blastogenesis and PHA reactivity in peripheral lymphocyte may be one of the useful methods for early prediction of rejection episode in lung transplantation.     

Reactive lymphocyte blastogenesis in canine renal transplantation.

Reactive lymphocyte blastogenesis was investigated by 3H-thymidine uptake of peripheral lymphocytes. Canine renal allotransplantations were performed in 19 dogs, which were divided into three groups by different immunosuppressive therapies. The first group, not treated with any immunosuppressants, showed the highest 3H-thymidine uptake, but the third group, treated with prednisolone (1.5 mg/kg/day) and azathioprine (4 mg/kg/day), did not show remarkable 3H-thymidine uptake except for 2 dogs which died of severe acute rejection. Single use of prednisolone (second group) was less effective in suppression of blastogenesis compared with the combined use of azathioprine and prednisolone. In addition to these experiments, the effects of surgical operation, blood transfusion, and infection on lymphocyte blastogenesis were investigated. Renal autotransplantation did not produce remarkable blastogenesis, but blood transfusion and infection caused strong lymphocyte blastogenesis. Periodical increases of 3H-thymidine uptake were observed in the infected animal.     

Hormonal therapy and alteration of lymphocyte proliferation.

Suppression of the blastogenic response of phytohemagglutinin-stimulated peripheral blood lymphocytes in autologous serum obtained from patients with prostatic cancer. Peyronie's disease, and transsexuals following estrogen therapy has been observed. Observations of suppression of blastogenesis in patients with prostatic cancer and without malignancy receiving estrogen suggests that this suppression would appear not to be associated with a particular pathologic state or malignancy per se but, rather, the given mode of therapy. The relevance of these observations to the effect of estrogen on cell-mediated responsiveness in patients with hormonally dependent tumors and to the suggested association between uterine cancer and prolonged administration of diethylstilbesterol and the development of vaginal tumors in offspring found in association with material ingestion during pregnancy remains speculative.     

Comparison of ketoconazole and amphotericin B in interference with thymidine uptake by and blastogenesis of lymphocytes stimulated with Histoplasma capsulatum antigens.

Human peripheral blood lymphocytes stimulated with Histoplasma capsulatum yeasts were exposed in culture to graded concentrations of ketoconazole or amphotericin B and subsequently assessed for membrane integrity, thymidine uptake, and blastogenesis. Lymphocyte reactivity varied with concentration and duration of exposure to ketoconazole. Overt membrane toxicity resulted from exposure to 40 micrograms of ketoconazole per ml for 5 days, diminished thymidine uptake occurred with concentrations as low as 5 micrograms/ml, and 15 to 20 micrograms/ml caused a marked decrease in thymidine uptake and eventually diminished blastogenesis. The antilymphocyte action of ketoconazole was neutralized by increasing the concentration of human serum in cultures to 40% regardless of its cholesterol content. Amphotericin B activity was qualitatively similar but less pronounced.     

Enhancement of Human Lymphocyte Transformation by Aggregated Human Gamma Globulin

The effect of heat-aggregated human gamma globulin (aggFII) on the induction of in vitro lymphocyte transformation, measured by the uptake of tritiated thymidine into newly synthesized DNA, was studied with peripheral blood lymphocytes derived from 12 patients with rheumatoid arthritis (RA), six with ankylosing spondylitis (AS), two with systemic lupus erythematosus (SLE), and seven normal subjects. It was found that 200 mug aggFII induced significant transformation of the lymphocytes of eight patients with RA, five with AS, one with SLE, and one normal subject. Neither deaggregated FII nor heat-aggregated human serum albumin induced significant transformation of the lymphocytes of any subject tested. A source of complement appeared necessary to support aggFII-induced blastogenesis, since enhanced transformation occurred only in the presence of fresh plasma. Heat-inactivated plasma and fetal calf serum (FCS), and FCS devoid of hemolytic complement, failed to support enhanced blastogenesis in the presence of aggFII. Since substrates similar to those employed in these studies are present in vivo in the rheumatoid joint, it is suggested that aggFII may enhance intra-articular lymphocyte transformation in subjects with RA.     

Therapeutic role of pericardiocentesis for acute necrotizing eosinophilic myocarditis with cardiac tamponade

We describe a patient with acute necrotizing eosinophilic myocarditis who recovered rapidly after pericardial drainage and without corticosteroid therapy. The 25-year- old man was referred to our hospital with suspected acute myocardial infarction on the basis of severe epigastralgia, abnormal Q waves and ST elevation on electrocardiography, and an increase in cardiac enzymes. Echocardiography disclosed pericardial effusion that compressed the right ventricle, left ventricular dysfunction in conjunction with posterolateral hypokinesis, and a thickened ventricular wall but no mural thrombus. The eosinophil count in the peripheral blood was slightly increased. Coronary angiography showed normal arteries and thus prompted an endomyocardial biopsy. The patient was transferred to the intensive care unit with a clinical diagnosis of myocarditis associated with cardiac tamponade. Emergency pericardiocentesis relieved symptoms immediately. The cells in the pericardial effusion were mainly eosinophils; interleukin 5 and interleukin 13 levels were predominantly elevated, and the effusion was drained for 5 days. The biopsy specimen revealed necrotizing eosinophilic myocarditis. Left ventricular function recovered within a week without corticosteroid therapy. No relapse was observed as of 8 months after diagnosis.     

In vitro proliferative response of chronic lymphocytic leukemia

Summary The peripheral blood lymphocytes of 13 previously untreated chronic lymphocytic leukemia (CLL) patients showed a decreased and delayed responsein vitro to plant mitogens (PHA and PWM) and specific antigens (PPD and MLC). In addition, the serum of these patients inhibited the mitotic reactivity of both autologous and homologous normal lymphocytes. Since incubation with CLL serum did not affect the SRBC-rosetting capacity of normal lymphocytes, we believe that CLL serum interferes with some metabolic stage in blastogenesis rather than at the level of mitogen membrane interaction. Thein vitro transformation of lymph node, bone marrow and peripheral blood lymphocytes in some of these patients was also investigated. Stimulation of peripheral blood cells with plant mitogens resulted in a more serious impairment of the response than that found with bone marrow and lymph node cells. This could be explained by the fact that, although CLL is a widespread lymphoproliferative disorder, some preferential homing of normally reactive cells in the bone marrow and lymph nodes cannot be excluded. Finally, since no stimulation was observed in mixed leukocyte cultures (MLC), our experiments provide evidence that no antigenic differences are detectable in CLL lymphoid populations.     

Defective leukocyte function-related cryoprecipitable protein in Cryoglobulinemia

Cryoprecipitable proteins (CPP), purified from plasmas removed during plasmapheresis treatment of patients with type II and type III cryoglobulinemia (CG), were evaluated in vitro for their effect on normal leukocyte function. CPP reduced normal mononuclear cell blastogenesis and polymorphonuclear cell phagocytosis responses in a dose-dependent fashion. At the maximum concentration studied, 5,000 (μg/ml, CPP were shown to suppress normal blastogenesis and phagocytosis 30–60% and 50–80% below the albumin control, respectively. A 50% suppression of normal blastogenesis and phagocytosis was observed at 500 p.g/ml for type III CPP. Leukocyte functions in CG patients were evaluated and compared to those of healthy controls. Blastogenesis and phagocytosis were significantly inhibited in CG patients. These studies suggest that CG patients have defective leukocyte function and that CPP have suppressive effects on normal leukocyte function. Our studies suggest that removal of CPP in CG patients may prove beneficial not only in reducing typical symptoms of CG but also in reducing the potencial inhibitory effects of CPP on leukocyte function.     

多发性硬化患者输注自体造血干细胞的护理与安全管理

目的探讨多发性硬化患者输注自体造血干细胞的护理措施与安全管理方法。方法回顾性分析2007年8月至2010年6月我科收治的8例多发性硬化患者输注自体造血干细胞的护理措施与安全管理。结果经全环境保护,心理护理,造血干细胞输注前、中、后护理,并发症防治等护理措施与安全管理,8例患者无外源性感染、出血等严重并发症发生,临床症状明显改善。结论精心的护理措施与安全管理能进一步保障多发性硬化患者输注自体造血干细胞的临床疗效。     

对急性呼吸系统感染患儿血清CK-MB增高的再认识

目的　探讨急性呼吸系统感染患儿血清CK MB增高而无心肌炎临床症状和心电图改变的原因。方法　对急性呼吸系感染患儿于病程的 1～ 5天内采静脉血同时测定CK MB(采用免疫抑制法 )和CTnⅠ (采用夹心ELISA法 ) ,并动态观察病人临床症状、心电图改变以判断是否合并心肌炎。结果　 13 0例急性呼吸系感染患儿中血清CK MB增高 5 8例 ,血清CTnⅠ增高 3例 ,其阳性率为 10 0 % ,经卡方检验P <0 .0 5 ,差异有显著性。 12 7例临床无心肌炎的病例中 ,血清CK MB增高 5 5例 ,正常 72例 ,12 7例患儿中血清CK MB增高组、CK MB正常组与正常儿童对照组三组间CTnⅠ值经方差分析P >0 .0 5 ,差异无显著性。结论　应用免疫法测定的血清CK MB值不能作为判断呼吸系感染时有无心肌损伤的“金指标” ,应改用质量法测定血清CK MB ,或测定血清CTnⅠ来补充。     

Stimulation by thymopoietin oligopeptides of lectin-dependent cell-mediated cytotoxicity in patients with systemic lupus erythematosus

The effect of thymopoietin penta- (TP-5), tetra-(TP-4), and tripeptides (TP-3) was studied on the depressed lectin-dependent cell-mediated cytotoxicity (LDCC) against adherent HEp-2 target cells by peripheral blood mononuclear cells from patients with active systemic lupus erythematosus (SLE). LDCC activity was evaluated by detachment from the monolayer of 3H-thymidine-prelabelled HEp-2 cells in the presence of concanavalin A (Con A). While 10(-5)M TP-3 moderated the depression, 10(-5) M TP-5 strongly enhanced LDCC activity in SLE patients up to the normal level. On the other hand, LDCC activity by normal donors was not influenced by TP-3 and TP-5. TP-4 had no major effect either in control or in SLE patients. In parallel experiments none of the thymopoietin peptides affected the Con A-induced suppressor activity on the blastogenesis of lymphocytes. A selective immunostimulatory effect of TP-3 and TP-5 on the generation of LDCC effector cells in patients with SLE is suggested.     

α-干扰素联合磷酸肌酸钠治疗病毒性心肌炎疗效分析

目的 探讨α-干扰素联合磷酸肌酸钠对病毒性心肌炎的临床疗效,为临床治疗病毒性心肌炎选择治疗药物提供依据.方法 选择2007年1月至2009年12月3年收住的23例病毒性心肌炎患者,随机将患者分为治疗组和对照组,两组均采取基础治疗及对症处理.治疗组在基础治疗的基础上使用α-干扰素联合磷酸肌酸钠治疗并观察4周;对照组在基础治疗的基础上加用辅酶Q10和病毒唑治疗,疗程相同.4周后对患者的临床症状和体征、辅助检查及临床疗效等情况进行分析比较.结果 治疗组在临床症状和体征的改善、辅助检查变化等方面均优于对照组,两组总有效率比较差异有统计学意义(P＜0.05).结论 α-干扰素联合磷酸肌酸钠治疗病毒性心肌炎有较好的疗效.     

Defective interleukin 2 production and responsiveness in human pulmonary tuberculosis

Patients with newly diagnosed, pulmonary tuberculosis had a tuberculin-specific defect in IL-2 production. Mean PPD-induced IL-2 activity was 81.2% lower in patients as compared with healthy tuberculin reactors. PPD-induced expression of T cell IL-2 receptors was 5.9 times less in peripheral blood mononuclear cells of patients with tuberculosis as compared with healthy tuberculin reactors. Furthermore, purified IL-2 failed to correct PPD-induced blastogenesis in patients. Suppression by adherent cells was operative in one group of patients; adherent cell depletion increased their T cell production of IL-2 7.2-fold. A second group of patients with low IL-2 production did not have suppressor adherent cells and were clinically distinct, with more extensive disease on chest x ray. The basis for low IL-2 production in such individuals is unknown. Disordered regulation of IL-2 metabolism may be a key feature in the depressed cellular immune response of tuberculosis.     

Determination of the circulating antibodies to cardiomyocyte membrane proteins based on the immunoblotting principle]

Cardiomyocyte membrane proteins (CMP) were isolated from the hearts of subjects dead from injuries or brain tumors by differentiated centrifugation in sucrose density gradient for the determination of anti-CMP antibodies in the blood serum samples of myocarditis patients. CMP preparation reactivity with blood serum antibodies of patients with myocarditis and other myocardial diseases, as well as with systemic lupus erythematosus, was assessed by the immune blotting technique. The findings evidence a high incidence of antibody interaction with CMP with a molecular mass of 67 kD. The method is highly specific and sensitive, its results are fairly well reproducible, and the technique is simple.     

Characterising vancomycin's immunotoxic profile using Swiss and CFW mice as an experimental model

Immunotoxicology can lead to determining the adverse effects of different compounds on the immune system. Sometimes, many drugs (especially antibiotics) induce immune alterations, mainly auto-immunity. This study was aimed at determining vancomycin's immunotoxic effect by comparing the original molecule to two of the most used copies. Thirty-two mice from two murine strains (Swiss and CFW) were treated with three antibiotic formulations for studying its effect on splenic lymphoid and peripheral blood cell populations by using haemograms, flow cytometry and blastogenesis assays. The results indicated that vancomycin produces neutropenia and lymphocytosis in peripheral populations and that it induces a selective immunomodulatory effect on splenocyte sub-populations, depending on formulation and the strain so treated.     

17 beta-carboxamide steroids: highly effective inhibitors of the phytohaemagglutinin mediated blastogenesis of normal human peripheral lymphocytes

Several novel 17 beta-carboxamide analogues of dexamethasone were synthesized. The common precursor, 9-fluoro-16 alpha-methyl-11 beta,17-dihydroxy-3-oxo-1,4-androstadiene-17 beta-carboxylic acid, did not bind to the glucocorticoid receptors of rat liver and human spleen tumours. In addition, no inhibition of the mitogen-induced blastogenesis of cultured human peripheral lymphocytes was observed. The 17 beta-carboxamide analogues, however, bound with similar affinities to the glucocorticoid receptors of both tissues. They inhibited the mitogen-induced blastogenesis of peripheral lymphocytes, showing the same potency and same order of binding affinity as the natural glucocorticoids.     

Diagnostic significance of various symptoms of right-ventricular insufficiency in patients with chronic obstructive bronchitis

The paper is concerned with the results of echocardiography, Doppler cardiography, radionuclide investigation and direct pressure measurement in the peripheral vein in 22 patients with chronic obstructive bronchitis and 7 persons with normal bronchial permeability. It was shown that the diameter of the peripheral veins, the pressure and rate of the blood flow in them and liver enlargement in patients with chronic obstructive bronchitis depended, to a large extent, on respiratory fluctuations of intrathoracic pressure. A conclusion was made that peripheral vein swelling and liver enlargement which were considered to be common symptoms of the development of right ventricular insufficiency, resulted from an increase in intrathoracic pressure during exhalation making difficult blood venous return to the heart in patients with dramatic disorders of bronchial permeability.