还原型谷胱甘肽抗白血病免疫佐剂作用实验研究

本研究探讨还原型谷胱甘肽(GSH)逆转反应性氧代谢物(ROM)对NK细胞抗白血病效应的抑制作用。在K562细胞、NK细胞的混合培养体系中分别加入富含单核细胞的单个核细胞(Mo)和白细胞介素-2(IL-2),观察ROM产量和K562细胞抑制率,然后分别加入GSH或二氢氯化物组胺(组胺),观察ROM产量及K562细胞抑制率的变化。结果表明:加入IL-2后,ROM的产量从33.17±25.02U/L增至223.59±59.41U/L(P<0.01),K562细胞抑制率从65.56%升至85.89%(P<0.01);在加入E/Mo=10/1、10/5、10/103种浓度的Mo后,ROM产量分别为389.79±43.83U/L,456.74±42.77U/L,601.42±21.92U/L,K562细胞抑制率分别为82.36%,81.36%,48.09%,加入组胺或GSH后,E/Mo=10/2时,ROM产量从389.79±43.83U/L,分别减至50.21±22.4U/L和-3.58±9.49U/L(P<0.05),随着GSH或组胺浓度的增加ROM产量逐减少,K562细胞抑制率从82.53%分别升至94.64%和96·39(P<0·05),ROM产量与K562细胞抑制呈负相关(P<0.05);E/Mo=10/5或10/10时,高浓度的GSH或组胺可使ROM产量减少,但K562细胞抑制率提高不明显(P>0.05)。结论:当E/Mo=10/2时,GSH逆转ROM强于组胺,提高NK细胞对K562的抑制率与组胺相似,但毒副作用轻微,GSH可能成为更理想的抗白血病的免疫佐剂。     

Unbound immunoglobulins are a major source of error in the quantitation of platelet surface-bound immunoglobulin levels

Analysis of assays for the determination of platelet-associated immunoglobulins (PAIgs) has led to disagreement over the amount of surface-bound Igs in both normal controls and patients with elevated platelet Ig levels. By the use of radiolabeled platelets and platelet counts, it was demonstrated that more than 10(8) platelets are disrupted after each centrifugation during platelet isolation procedures, releasing intraplatelet contents into the fluid phase of resuspended platelets in buffer. It was shown that suspensions of whole washed platelets contain a significant, but generally overlooked, amount of unbound Igs that have been liberated from platelets disrupted during processing. When platelet suspensions are evaluated for Igs with assays that fail to incorporate a final separation of whole platelets from the suspension fluid, unbound Igs as well as those bound to the platelet surface are measured, which yields a logarithmic overestimation of surface-bound Igs. It has been further demonstrated that patients infected with human immunodeficiency virus type 1 can have elevations of PAIgG, PAIgA, and PAIgM in both the thrombocytopenic and nonthrombocytopenic states. These elevations are due to increased internal platelet pools of Igs and not to increased surface-bound Igs.     

Specific insulin assays, insulin sensitivity and blood pressure

Serum insulin concentrations have been used as markers of insulin resistance in population studies examining the relationship between insulin resistance and blood pressure, but the relationship is variable among studies. We hypothesized that differences in cross-reactivity of insulin assays with proinsulin and its split/des-amino products might account for the variation. We therefore examined fasting and post- glucose load serum insulin concentrations (determined by both specific and conventional assays), insulin sensitivity (measured by the euglycaemic clamp technique), and blood pressure, in a group of 56 diabetic (NIDDM) and non-diabetic subjects. Insulin concentrations as measured by the two methods were highly correlated (r = 0.97, p < 0.0001), and the relationships among serum insulin concentrations, insulin sensitivity and blood pressure were independent of assay method; for example, in non-diabetic subjects the univariate correlation between log10AUC insulin and insulin sensitivity index was similar with both methods [r = -0.81 vs. r = -0.82, p < 0.0001 (specific vs. conventional assay)]. Discrepancies between studies in the relationship between serum insulin concentrations and blood pressure are unlikely to be due to cross-reactivity of conventional insulin assays with proinsulin-like molecules.      

Immune response gene function correlates with the expression of an Ia antigen. II. A quantitative deficiency in A(e):E(a), complex expression causes a corresponding defect in antigen-presenting cell function

A series of experiments were performed to explore the role of complementing major histocompatability complex (MHC)-linked immune response Ir genes in the murine T cell proliferative response to the globular protein antigen pigeon cytochrome c. The functional equivalence of I-E-subregion-encoded, structurally homologous E(a) chains from different haplotypes bearing the serologic specificity Ia.7 was demonstrated by the complementation for high responsiveness to pigeon cytochrome c of F(1) hybrids between low responder B 10.A(4R) (I-A (k)) or B 10.S (I-A(8)) mice and four low responder E(a)- bearing haplotypes. Moreover, this Ir gene function correlated directly with both the ability of antigen-pulsed spleen cells from these same F(1) strains to stimulate pigeon cytochrome c-primed T cells from B10.A or B10.S(9R) mice, and with the cell surface expression of the two-chain Ia antigenic complex, A(e):E(a), bearing the conformational or combinatorial determinant recognized by the monoclonal anti-Ia antibody, Y-17. The B 10.PL strain (H-2(u)), which expresses an Ia.7-positive I-E- subregion-encoded E(a) chain, failed to complement with B10.A(4R) or B10.S mice in the response to pigeon cytochrome c. However, (B10.A(4R) × B10.PL)F(1) and (B10.S × B10.PL)F(1) mice do express A(k)(e):E(u)(a) and A(8)(e):E(u)(a) on their cell surface, although in reduced amounts relative to A(k,s)(e):E(k,d,p,r)(a) complexes found in corresponding F(1) strains. This quantitative difference in Ia antigen expression correlated with a difference in the ability to present pigeon cytochrome c to B 10.A and B 10.S(9R) long-term T cell lines. Thus, (B10.A(4R) × B10.PL)F(1) spleen cells required a 10-fold higher antigen dose to induce the same stimulation as (B10.A(4R) × B10.D2)F(1) spleen cells. In addition, the monoclonal antibody, Y-17, which reacts with A(e):E(a) molecules of several strains, had a greater inhibitory effect on the proliferative response to pigeon cytochrome c of B10.A T cells in the presence of (B10.A(4R) X B10.PL)F(1) spleen cells than in the presence of (B10.A(4R) X B10.D2)F(1) spleen cells. These functional data, in concert with the biochemical and serological data in the accompanying report, are consistent with the molecular model for Ir gene complementation in which appropriate two-chain Ia molecules function at the antigen-presenting cell (APC) surface as restriction elements. Moreover, they clearly demonstrate that the magnitude of the T cell proliferative response is a function of both the concentration of nominal antigen and of the amount of Ia antigen expressed on the APC. Finally, the direct correlation of a quantitative deficiency in cell surface expression of an Ia antigen with a corresponding relative defect in antigen-presenting function provides strong independent evidence that the I-region-encoded Ia antigens are the products of the MHC-linked Ir genes.     

Anti-Ata causing mild hemolytic disease of the newborn

We report here a case of moderately severe hemolytic disease of the newborn (HDN) due to anti-Ata. The gravida 5 proposita was group A rr and previously was found to have anti-Ata and -D. At the 35-week mark of this pregnancy, her anti-Ata demonstrated a titer of 256, score 79. Fluid obtained by amniocentesis at 36 weeks showed an optical density at 450 nm of 0.08 (midzone). The baby was delivered at 38 weeks by cesarean section. The cord cells were group A rr with a 3+ direct antiglobulin test. The dichloromethane eluate of the infant's cells demonstrated anti-Ata specificity only. At birth, the infant's total bilirubin (TB) was 2.1 mg per dl and the hematocrit level (Hct) was 33.8 percent. Within 8 hours, the TB had risen to 3.8 mg per dl. Phototherapy was initiated at 7-1/2 hours and maintained for 40 hours. The infant's TB rose to a maximum level of 10.5 mg per dl 24 hours after phototherapy was discontinued. At discharge 4 days postpartum, the infant's TB had dropped to 9.2 mg per dl, and the Hct value was 38 percent. On a visit 7 days postpartum, the infant's TB level had fallen to 6.5 mg per dl and the hct value was 38 percent. Transfusions were not necessary.     

Major histocompatibility complex markers and red cell antibodies to the Rh (D) antigen. Absence of association

Between 20 and 35 percent of Rh(D) antigen-negative individuals do not develop antibodies to D even after multiple transfusions of Rh-positive red cells. To evaluate the possibility that antibody production after exposure to the D antigen was related to a major histocompatibility complex immune response gene, analysis of the HLA genotypes of 38 Rh-sensitized women and their families was performed. No significant deviations were found in the frequency of any individual HLA class I, II, or III allele or of any extended haplotype (fixed allelic combinations of HLA-B, HLA-DR, and the complement components BF, C2, C4A, and C4B). Type 1 errors due to the extreme allelic polymorphism of the HLA system, as well as the ethnic variation in patient groups, may have contributed to HLA allele-antibody responder relationships reported in earlier studies.     

高压氧治疗弥漫性轴索损伤的临床疗效观察

目的：探讨高压氧治疗弥漫性轴索损伤的疗效。方法：60例弥漫性轴索损伤患者分为治疗组32例和对照组28例，观察GCS评分平均值的变化和GOS预后情况。结果：治疗组GCS评分平均值曲线抬高，病死率下降，GOS评分高于对照组，其差异有显著性意义（P〈0.05）。结论：高压氧辅助治疗弥漫性轴索损伤可以明显降低病死率，改善患者的预后     

Transient neutropenia induced by transfusion of blood exposed to nylon fiber filters

During the course of granulocyte collection by continuous-flow filtration leukopheresis, an abrupt fall in neutrophil count was noted (mean decrease 77%, range 64%-95%). Neutropenia occurred within 5 min of return of blood exposed to the nylon fiber filters and lasted less than 30 min. Saline exposed to the fibers, withdrawal and reinfusion of whole blood, and heparin did not cause neutropenia. Heparinized blood passed by gravity through isolated filters and reinfused immediately also induced neutropenia (mean decrease 64% +/- 8%, range 11%-19%). Blood anticoagulated with ACD (decrease 19.5% +/- 6%, range 6%-56%), heparinized plasma (N = 10, decrease 15% +/- 3%, range 3%-29%) and platelet-rich plasma exposed to the filters failed to produce neutropenia. 91% +/- 2% of the neutrophils adhered to the fibers using heparinized blood as compared to 21% +/- 5% using ACD (p less than 0.001). All donors were asymptomatic during the infusions. These results suggest that during neutrophil adherence a substance is released which produces profound, transient neutropenia perhaps by inducing margination of cells.     

Prolonged administration of granulocyte colony-stimulating factor (filgrastim) to patients with Fanconi anemia: a pilot study

This report examines the effect of filgrastim (granulocyte colony- stimulating factor, [G-CSF] in 12 patients with neutropenia [absolute neutrophil count [ANC] < 1,000/mm3]) caused by Fanconi anemia (FA). Two of 14 patients who were evaluated for study entry were ineligible because of unsuspected cytogenetic abnormalities in their bone marrow (BM). G-CSF was started at 5 micrograms/kg/d. All patients had an increase in their ANC at week 8 (mean increase = 15,664/mm3). The median ANC during therapy was 5,030/mm3. Eight of 10 patients who completed 40 weeks on study maintained an ANC > 1,500/mm3 on G-CSF given every-otherday. Four patients had an increase in their platelet count by week 8 without transfusion (maximum increase = 23,000 to 45,000/mm3); however, platelet counts fell toward baseline levels as the G-CSF dose was reduced. BM CFU-MK were increased at week 8 in three of four evaluable patients. Four patients who did not receive red blood cell transfusions had an increase in their hemoglobin level of at least 2.0 g/dL. A fifth patient had a red blood cell transfusion in week 2 and then had a similar increase in hemoglobin level without subsequent transfusion. Eight of 10 patients who completed 40 weeks of treatment showed increases in the percentage of BM CD34+ cells measured by flow cytometry. The same proportion showed increases in peripheral blood CD34+ cells. Increased BM cellularity and myeloid hyperplasia were constant findings and were associated with increased expression of the proliferating cell nuclear antigen. Adverse experiences were mild fever (1 patient) and a new BM cytogenetic abnormality at week 40 (1 patient). This study shows that prolonged administration of G-CSF exerts a stimulatory effect on the BM of FA patients and may be used to maintain a clinically adequate ANC in these patients. G-CSF has beneficial effects on multiple hematopoietic lineages in some patients and may be a good candidate for use in combination cytokine protocols for FA patients with progressive aplastic anemia. G-CSF use results in an increase in circulating CD34+ cells, a finding with important implications for future gene transfer protocols.