Immunologic mechanisms in hypersensitivity pneumonitis: I. Evidence for cell-mediated immunity and complement fixation in pigeon breeders' disease

Immunologic studies were performed on 5 patients with pigeon breeders' disease. Intradermal injection of pigeon serum produced an immediate wheal-and-flare reaction within 15 minutes and a secondary Arthus-type reaction within 4 to 8 hours. Immunofluorescent studies of the secondary reaction site showed IgG, C3, and C4 in 2 patients. Patients' sera produced multiple precipitin bands with pigeon serum when reacted by double diffusion in gel. IgG antibody isolated from each of the patients' serum formed precipitating immune complexes that fixed large amounts of complement (C4) when added to fresh human serum. Peripheral blood lymphocytes from 4 of the 5 patients produced macrophage migration inhibitory factor (MIF) when challenged with dilute pigeon serum. These studies are the first to show complement fixing antibodies and macrophage MIF production by lymphocytes from patients with hypersensitivity lung disease and suggest that both humoral and cellular immunity may be important in the pathogenesis of these disorders.     

Chronic myelogenous leukemia in the monosomic cell line of a fertile turner syndrome mosaic (45,X/46,XX)

Malignancy in patients with constitutional chromosome abnormality is of interest not only because it permits insights into the relationship between chromosome abnormality and cancer, but also because it provides opportunities to address such questions as the clonality and evolution of tumors. We report Ph¹-positive chronic myelogenous leukemia (CML) in a 50-year-old mosaic (45,X/46,XX) Turner syndrome patient whose leukemia was restricted to the monosomic cell line. Our extensive cytogenetic studies of this patient demonstrated that non-leukemic normal cells persisted in the marrow and were able to proliferate during a period of temporary suppression of the leukemic clone following aggressive treatment.     

人类白细胞抗原-G突变体cDNA克隆及在K562细胞上的表达

目的：克隆人类白细胞抗原-G(Human leukocyte antligen-G，HLA-G)突变体cDNA，并使其在HLA-I类阴性的K562细胞上获得稳定表达，为研究配-受体之间的识别机制奠定基础。方法：用RT-PCR方法从人子宫蜕膜组织扩增出HLA-GcDNA，得到全长HLA-GPCR产物后，用桥式PCR方法进行定点点突变，将突变的目的基因亚克隆于逆转录，将突变的目的基因亚克隆于逆转录mG-pLNCX表达载体，采用感染的方法将重组质粒转入K562细胞，最后经G418筛选及有限稀释，利用单克隆抗体W6/32进行FACS及mRNA检测，观察HLA-G突变体在靶细胞表面的表达。结果：HLA-G突变体分子在经mG-pLNCX转染的靶细胞表面获得稳定高表达。结论：成功构建了mG-pLNCX表达载体，并使HLA-G突变体分子在HLA-I类阴性的靶细胞K562细胞上获得稳定表达。     

Demonstration of an inwardly rectifying K+ current component modulated by thyrotropin-releasing hormone and caffeine in GH3 rat anterior pituitary cells

 Reduction of an inwardly rectifying K⁺ current by thyrotropin-releasing hormone (TRH) and caffeine has been considered to be an important determinant of electrical activity increases in GH₃ rat anterior pituitary cells. However, the existence of an inwardly rectifying K⁺ current component was recently regarded as a misidentification of an M-like outward current, proposed to be the TRH target in pituitary cells, including GH₃ cells. In this report, an inwardly rectifying component of K⁺ current is indeed demonstrated in perforated-patch voltage-clamped GH₃ cells. The degree of rectification varied from cell to cell, but both TRH and caffeine specifically blocked a fraction of current with strong rectification in the hyperpolarizing direction. Use of ramp pulses to continuously modify the membrane potential demonstrated a prominent blockade even in cells with no current reduction at voltages at which M-currents are active. Depolarization steps to positive voltages at the maximum of the inward current induced a caffeine-sensitive instantaneous outward current followed by a single exponential decay. The magnitude of this current was modified in a biphasic way according to the duration of the previous hyperpolarization step. The kinetic characteristics of the current are compatible with the possibility that removal from inactivation of a fast-inactivating delayed rectifier causes the hyperpolarization-induced current. Furthermore, the inwardly rectifying current was blocked by astemizole, a potent and selective inhibitor of human ether-á-go-go -related gene (HERG) K⁺ channels. Along with other pharmacological and kinetic evidence, this indicates that the secretagogue-regulated current is probably mediated by a HERG-like K⁺ channel. Addition of astemizole to current-clamped cells induced clear increases in the frequency of action potential production. Thus, an inwardly-rectifying K⁺ current and not an M-like outward current seems to be involved in TRH and caffeine modulation of electrical activity in GH₃ cells. Received: 15 May 1997 / Received after revision and accepted: 24 July 1997     

Functional mapping of the activity of the R region in the human T-cell leukemia virus type I long terminal repeat to increase gene expression

We previously demonstrated the activity of the R fragment in the long terminal repeat of human T-cell leukemia virus type I for elevation of the level of gene expression. In this study, the fragment was deleted with BAL31 nuclease to determine its functional domain. Series of the shortened R fragments were linked to the simian virus 40 promoter unit, which regulated expression of a reporter gene. Examination with the R fragments deleted from the 5 and 3 ends showed that borders of the functional domain were mapped within nucleotide positions 458 to 473 for the 5 end and nucleotide positions 559 to 594 for the 3 end, respectively. Thus we conclude that a 136-base-pair fragment corresponding to the second half of the R region was sufficient to allow elevation of the level of gene expression.     

胎肝细胞质液对重症病毒性肝炎患者外周血TL－CFU及mIL－2R影响的实验观察

采用半固体一步单层琼脂培养法和单克隆荧光抗体技术分别观察重症肝炎外周血ＴＬ－ＣＦＵ和ｍＩＬ－２Ｒ，发现重症肝炎患者ＴＬ－ＣＦＵ（１０４．４±３２．６）及ｍＩＬ－２Ｒ（３５．６±８．６）较正常人明显降低。在培养体系中加胎肝细胞质液后，无论在重症肝炎组还是在正常组均不能明显地提高ＴＬ－ＣＦＵ，说明胎肝细胞质液不含具生物佐的促ＴＬ－ＣＦＵ因子。但对ｍＩＬ－２Ｒ表达的影响，在重症肝炎组病人，只有在ＰＨＡ存在条件下才能促进ｍＩＬ－２Ｒ的表达，说明胎肝细胞质波含有某种（些）物质能协同ＰＨＡ促进重症肝炎患者外周血淋巴细胞ｍＩＬ－２Ｒ表达。     

Biochemical characterization of I-Ak proteins from mutant antigen-presenting B-cell--B-lymphoma hybridomas

Chemically induced mutants of an I-Ak,d expressing antigen-presenting B-cell--B-lymphoma hybridoma have recently been generated by immunoselection in vitro and were found to possess alterations in some of their serologically and functionally defined I-Ak region dependent functions. In order to identify at the structural level the origin of the differences in serological and functional properties of these mutants, I-Ak molecules from several of these mutant hybridomas were compared biochemically to wild-type I-Ak polypeptides by two-dimensional gel electrophoresis and high-pressure liquid chromatographic tryptic peptide analyses. Two-dimensional gel electrophoresis indicated that no major structural alterations, resulting in changes in mol. wt or charge, had occurred in the Ak alpha or Ak beta polypeptides from the mutant cells. Likewise, Ak alpha peptide maps of the mutants were indistinguishable from the normal Ak alpha peptide maps. However, two of the three mutants studied did exhibit one additional peptide in their Ak beta peptide maps. These results suggest that the major deficiencies in T-cell-activating functions of these mutants are a result of a limited alteration in the Ak beta polypeptide primary structure.     

Kinetics and distribution of voltage-gated Ca,Na and K channels on the somata of rat cerebellar Purkinje cells

Voltage gated ion channels on the somatic membrane of rat cerebellar Purkinje cells were studied in dissociated cell culture with the combination of cell-attached and whole-cell variation of patch clamp technique. The method enables us to record local somatic membrane current under an improved space clamp condition. Transient (fast-inactivating) and steady (slow inactivating) Ca channel currents, Na current, transient (fast-inactivating) and steady (slow-inactivating) K currents, were observed. Transient and steady Ca channel currents were activated at test potentials more positive than –40 mV and –20 mV, respectively (in 50 mM external Ba). The transient current inactivated with a half-decay time of 10–30 ms during maintained depolarizing pulses, while the steady current showed relatively little inactivation. Na current was activated at more positive potentials than –60 mV, and inactivated with a half-decay time of less than 5 ms. Transient and steady K outward currents were recorded at more positive potential than –20 mV and –40 mV, respectively. The transient current inactivated with a half-decay time of 2–8 ms. Ca, Na and K channels showed different patterns of distribution on the somatic membrane. Steady Ca channels tended to cluster compared with Na or K channels.     

普乐可复对难治性肾病综合征患者血清IL-2、sIL-2R的影响及其临床意义

目的:研究普乐可复(FK506)对难治性肾病综合征患者血清IL-2、sIL-2R的影响及其临床意义。方法:应用酶联免疫吸附法检测难治性肾病综合征患者经普乐可复治疗前后血清IL-2、sIL-2R水平变化,并监测患者24小时尿蛋白、血浆白蛋白、血脂的变化。结果:难治性肾病综合征患者经普乐可复治疗前血清IL-2、sIL-2R水平均显著高于正常对照组(P<0.05)。治疗后血清IL-2、sIL-2R水平较治疗前明显下降(P<0.05)。治疗前24小时尿蛋白水平显著高于正常对照组(P<0.01),血浆白蛋白显著低于正常对照组(P<0.01),血脂水平显著高于正常对照组(P<0.01);与治疗前比较,治疗后24小时尿蛋白水平显著下降(P<0.05),血浆白蛋白水平显著升高(P<0.01),血脂水平显著下降(P<0.05)。治疗后组与正常组比较,除IL-2外,余各项指标均无显著性差异(P>0.05)。结论:在难治性肾病综合征患者体内存在IL-2、sIL-2R的异常,普乐可复对其有明确的抑制作用,从而调节T细胞活性,有效降低24小时尿蛋白,提高血浆白蛋白含量,降血脂,缓解难治性肾病综合征的病情。     

奥美拉唑对家兔肾脏内髓集合管细胞H+/K+交换功能的影响

目的探讨奥美拉唑对家兔肾脏IMCD细胞H+/K+交换的影响,以及经洗涤处理是否解除这种影响.方法原代培养家兔肾脏IMCD细胞单层,在100 μmol/L奥美拉唑缓冲液中孵育25 min, 洗涤组则在奥美拉唑缓冲液孵育后,用不含奥美拉唑的缓冲液洗涤IMCD细胞;对照组在不含奥美拉唑的缓冲液孵育25 min;CECF/AM荧光探针法测定各组IMCD细胞H+/K+交换.结果奥美拉唑组的H+/K+交换为(0.016±0.006) dpHi/min(n=8);与对照组的(0.053±0.008) dpHi/min(n=8)相比,相差非常显著(P<0.001);洗涤组的H+/K+交换为(0.016±0.006) dpHi/min(n=6),与对照组(n=6)的(0.052±0.009)dpHi/min相比,相差非常显著(P<0.001).结论 100 mol/L的奥美拉唑对家兔IMCD细胞的H+/K+交换有显著影响,而且洗涤处理不能解除这种抑制.因而,肺心病合并上消化道出血患者使用奥美拉唑时,应综合考虑其利弊.