The use of human lymphoid cell line lymphokine preparations (LCL-LK) as working standards in the bioassay of human leucocyte migration inhibition factor (LIF)

The leucocyte migration inhibition test in agarose as described by Clausen (1971) was modified into a statistically designed assay of LIF activity using human polymorphonuclear leucocytes from single blood donors. Individual assays included a laboratory standard of lymphokine with LIF activity prepared from the culture supernatants of the RPMI 1788 human lymphoblastoid cell line (LCL-LK). Analysis of 157 LIF assays revealed simple criteria by which the acceptability of an individual assay could be judged before subjecting it to statistical analysis. The failure of LIF assays to meet these criteria of acceptability was particularly associated with low areas of control polymorph migration in the absence of lymphokine (‘spontaneous migration’).We demonstrate that the statistically designed assay permits the measurement, with precision, of LIF activity in units/ml by reference to a working standard of LCL-LK. We illustrate the use of this assay in the measurement of LIF activity generated by tuberculin-stimulated human peripheral blood lymphocytes.     

三物降压汤对高血压病患者血压及淋巴因子激活的杀伤细胞的影响

目的：研究三物降压汤对血压、淋巴因子激活的杀伤（ＬＡＫ）细胞的影响及其可能机制。方法：用随机、单盲、自身、组间平行对照法用三物降压汤对轻度高血压病患者（n＝３０）进行为期８周的治疗观察，并与开搏通治疗组（n＝３０）比较。分别检测治疗前后的血压、ＬＡＫ细胞的增殖与活性、ＬＡＫ细胞表达的超氧化物歧化酶（ＳＯＤ）样物质及其对自由基的清除能力，为进一步了解ＬＡＫ细胞与血压之间的关系，并观察了自发性高血压大鼠（ＳＨＲ）胸主动脉环对硝酸甘油和乙酰胆碱（Ａch）的舒张反应以及ＬＡＫ细胞与ＳＯＤ标准品对ＳＨＲ胸主动脉环对Ａch舒张反应的影响并与京都威斯特大鼠及经三物降压汤治疗后的ＳＨＲ比较。结果：三物降压汤在降血压同时，ＬＡＫ细胞增殖、活性、其所表达的ＳＯＤ及清除自由基效应均明显增加。结论：三物降压汤既能降血压又能调节免疫功能。     

Major histocompatibility complex (MHC) class II restriction,lymphokine production,and IgE regulation of house dust mite-specific T-cell clones

To elucidate the regulatory mechanism of human IgE synthesis, we have cloned house dust mite (Dermatophagoides pteronyssinus; Dp)-specific T-cell clones from three asthmatic children and three healthy individuals. Twelve clones were cloned from each group. All of these clones were CD3⁺, CD4⁺, CD8^–, and HLA-DR⁺. After stimulation with allergen in the presence of antigen presenting cells (APCs), half of the T-cell clones from asthmatic children and one-third of those from normals produced interleukin 4 (IL-4). None of the patients' clones produced interferon r (IFN-r), while 10 of 12 normals' clones did. After stimulation with calcium ionophore A23187 and phorbol myristic acetate (PMA), the production of IL-4 was markedly increased in both patients and normals. However, only 3 of the 12 patients' clones produced IFN-r, while all of the normals' clones did. The T-cell clones of both patients and normals produced comparable IL-2. To study the kinetics of lymphokine productions, a HLA-DRw12-restricted T-cell clone (FYD 3.1) was stimulated, respectively, with a combination of A23187 and PMA, phytohemagglutinin (PHA), or Dp antigen in the presence of APCs. Maximal IL-2 and IL-4 productions were detected 12 hr after A23187 and PMA stimulation, whereas IFN-r could not be detected even 36 hr after stimulation. When stimulated with PHA, the production of IFN-r peaked on the fourth day, but IL-4 was not detected. After stimulation with Dp antigen and APCs, IL-4 and IL-2 were detected on the second and third days, but IFN-r was not detected. The IgE production by autologous purified B cells in the presence of allergen or IL-4 was found to be augmented by the FYD 3.1 T-cell clones. IFN-r was observed to counteract the effects of the T-cell clones and IL-4. Thus, the secretory patterns of lymphokine and kinetics of lymphokine production of allergen-specific T-cell clones can be used to explore the regulatory mechanism of human IgE synthesis.     

Action of the human lymphokine histamine releasing factor on mouse peritoneal mast cells

Histamine releasing factor (HRF)--a human lymphokine--has been shown previously to release histamine from basophils in vitro. In this paper we show that HRF acted across the species barrier and released histamine from mouse peritoneal mast cells. This response was dose-dependent. Mast cells from both sensitized and non-sensitized mice were equally susceptible to the action of HRF. We observed synergistic action of HRF with specific allergen (ovalbumin) or HRF with anti-IgE antibody in releasing histamine from mast cells. Preincubation of mast cells with calcium ion chelating agent ethylenediaminetetraacetic acid (EDTA) or disodium cromoglycate induces only a small inhibition of histamine release caused by HRF. We conclude that histamine release from mouse peritoneal mast cells can serve as an in vitro test for the assay of human HRF.     

多疗程异体LAK/rIL-2治疗中晚期肝癌33例分析

研究了异体ＬＡＫ／ｒＩＬ－２治疗方案,经治的３３例晚期肝癌中,ＣＲ１例（３％）,ＰＲ１０例（３０％）,总有效率３３％,副作用小。提示该治疗方案对中晚期肝癌是一种可行的有效治疗。     

Immunotherapy of metastatic kidney cancer.

From April 1986 to September 2000, 122 MRCC patients were treated by monthly intralymphatic injections (containing a mean of 573 IL-2 U and 26 x 10(6) LAK cells) and i.m. administration of IFN and TF; 71 patients also received a 3-day cycle of monthly IL-2 inhalations with a mean of 998 daily U. MRCC cases not treated by immunotherapy (n = 89) represent our historical controls. Adverse clinical side effects related to treatment were negligible. CR (n = 11) and PR (n = 13) were noticed in 24/122 patients. Of 24 responding patients, 17 resumed progression, whereas 7 remain in remission 11-69 months later. The overall median survival of treated patients (28 months) was 3.5-fold higher than the median survival of historical controls (7.5 months), and a Kaplan-Meier curve showed 25% survival 11 years after the beginning of immunotherapy. Apparently, the addition of IL-2 by inhalation improved survival. The present immunotherapy protocol appears to be efficacious, safe, devoid of adverse side effects, far less costly than others and able to offer a good quality of life to MRCC patients; if confirmed in a multicenter trial, it could set the basis for developing low-dose immunomodulatory treatments.     

Evidence for a Low-Affinity Interleukin-3 Receptor

Abstract

Interleukin-3 (IL-3) regulates the proliferation of myeloid, erythroid, and lymphoid cells. Previous reports showed IL-3 binding restricted to a single high-affinity (K_d = 50-200 pin) site. Here, we demonstrate by equilibrium studies an additional binding site for IL-3 with lower apparent affinity (K_d=5-20 iym). Furthermore, kinetic analysis shows that two binding sites for IL-3 exist: IL-3 dissociates slowly from the first site (I_½=4hr; k_?1=2.7x10^?3 min^?1), whereas it dissociates rapidly (T_½=4.0 min; k_?1 = 0.116 min^?1) from the second site. Cross-linking showed that [¹²⁵I]IL-3 binding to the 115- and 140-kD proteins was not saturable at concentrations commensurate with high-affinity binding and IL-3 dissociated rapidly from these same molecules. Thus, the low affinity IL-3 receptor is a molecule(s) of 115- to 140-kD.     

CD_3AK细胞的培养与抗肿瘤活性研究

目的体外扩增经ＣＤ３单抗（ＣＤ３ＭｃＡｂ）活化的杀伤细胞（ＣＤ３ＡＫ），并测定其抗肿瘤细胞作用和表型变化。②方法取１３例肺癌和２例肝癌病人外周血，经密度梯度离心获取淋巴细胞，以ＣＤ３ＭｃＡｂ和ＩＬ－２作为诱导剂，采用液相三步扩增方法扩增培养，并分别用ＭＴＴ和ＩＦＡ法与自体ＬＡＫ对照细胞比较细胞动力学、细胞毒作用和表型变化。③结果ＣＤ３ＡＫ细胞扩增速度、数量和抗肿瘤活性均优于自体ＬＡＫ细胞，其表型为ＣＤ３，ＣＤ４，ＣＤ８和ＣＤ１６的特殊异源淋巴细胞群。其中ＣＤ４亚群表型数量随培养时间延长明显升高（χ２＝７．６６，Ｐ＜０．０１），而ＣＤ１６则明显下降（χ２＝５．２３，Ｐ＜０．０５），该亚群表型变化可能与培养时间有关。④结论ＣＤ３ＡＫ细胞具有较强的杀肿瘤细胞作用，同时也可作为基因治疗的理想载体细胞。     

Therapy of recurrent high grade gliomas with surgery,and autologous mitogen activated IL-2 stimulated killer (MAK) Lymphocytes: I. Enhancement of MAK lytic activity and cytokine production by PHA and clinical use of PHA

Nineteen patients with recurrent high grade gliomas were treated in a phase I/II trial with aggressive debulking of the tumor, mitogen activated IL-2 stimulated peripheral blood lymphocytes (MAK cells), and rIL-2. Phytohemagglutin (PHA) was introduced into the tumor site in 16 patients prior to implanting MAK cells and IL-2 in an attempt to trigger more effective lysis of the tumorin vivo. In vitro both TNF bioactivity and cytolytic activity of long term cultured MAK (LMAK) cells were dramatically enhanced by adding PHA to the cultures of these activated PBL. Three of eleven patients (27%) had a decrease in size of the enhancing lesion on CT and/or MRI. Seven (37%) patients clinically improved. Median survival after therapy was 30 weeks. PHA was shown to be safein vivo and more effective than IL-2 triggering enhanced effector functionin vitro.     

妇痛宁联合免疫法对子宫内膜异位症免疫功能的影响

目的:探讨中药妇痛宁煎剂联合过继免疫疗法(AdoptiveimmunotherapyAIT)对子宫内膜异位症免疫功能的影响。方法:50只S—D大鼠随机分为空白对照组、模型对照组、妇痛宁煎剂灌胃组、LAK细胞注射组、妇痛宁煎剂灌胃联合LAK细胞注射共5组。分别测定各组大鼠腹腔中自然杀伤细胞(NK细胞)活性,血清中白细胞介素—1(IL—1)、及血清E2、P、PRL的水平。结果:3个治疗组腹腔液中NK细胞活性均有不同程度提高,血清IL—1水平均有所降低,且都以联合治疗组数值变化显著;血清E2、P水平妇痛宁组及联合组显著下降,LAK细胞组无明显改变。结论:妇痛宁联合过继免疫法治疗子宫内膜异位症较单独应用妇痛宁或过继免疫疗法更有效地调节内异症紊乱的免疫机制。提示中西医结合治疗子宫内膜异位症具有诱人前景。