经支气管动脉输液LAK／IL—2治疗肺癌的免疫指标变化及临床意义

５４例原发性肺癌患者轻支气管动脉输注ＬＡＫ／ＩＬ－２前后分别测定了体内ＮＫ活性，淋转，Ｔ细胞亚群及体液免疫指标的变化，结果表明，治疗前体内ＮＫ，淋转，ＯＫＴ３，ＯＫＴ４均低于正常值，ＯＫＴ８，Ｃ３，ＩｇＡ，ＩｇＭ治疗前均高于正常值，治疗后大部分观察指标趋于正常。     

天黄汤对鼻咽癌放疗患者免疫功能的影响

作者采用随机对照分组的方法，观察了中药天黄汤对鼻咽癌放疗患者免疫功能的影响。３６例患者分为两组：（１）仅用放射治疗（１８例）；（２）放疗加天黄汤治疗（１８例）。免疫指标包括：ＩｇＧ、ＩｇＡ、ＩｇＭ、补体Ｃ３、淋巴细胞百分数（Ｉｇ）、ＡＮＡＥ阳性率，ＮＫ细胞活性。结果发现，与第一组相比较，第二组患者ＩｇＧ、ＩｇＡ水平下降，淋巴细胞百分数、ＡＮＡＥ阳性率、ＮＫ细胞活性升高。经统计学处理，有显著性差异（     

THE IN VITRO POTENTIATION OF LAK CELL CYTOTOXICITY IN CANCER AND AIDS PATIENTS INDUCED BY F3—A FRACTIONATED EXTRACT OF ASTRAG

ＴＨＥＩＮＶＩＴＲＯＰＯＴＥＮＴＩＡＴＩＯＮＯＦＬＡＫＣＥＬＬＣＹＴＯＴＯＸＩＣＩＴＹＩＮＣＡＮＣＥＲＡＮＤＡＩＤＳＰＡＴＩＥＮＴＳＩＮＤＵＣＥＤＢＹＦ３—ＡＦＲＡＣＴＩＯＮＡＴＥＤＥＸＴＲＡＣＴＯＦＡＳＴＲＡＧＡＬＵＳＭＥＭＢＲＡＮＡＣＥＵＳＣｈｕ...     

成纤维细胞介导的人α型干扰素基因疗法与过继免疫化疗联合治疗人肝癌的实验研究

作者以皮下接种人肝细胞癌细胞ＳＭＭＣ７７２１的裸鼠为荷瘤模型，由人外周血单个核细胞经体外ＩＬ－２和丝裂霉素Ｃ灭活的人肝癌细胞ＳＭＭＣ７７２１共同刺激培养后激活的杀伤细胞（ＡＫ），研究了将成纤维细胞介导的人ＩＦＮ－α基因疗法与ＩＬ－２／ＡＫ／阿霉素过继免疫化疗联合应用后对肝癌的治疗作用。结果表明：（１）给肝癌裸鼠单独腹腔埋植人ＩＦＮ－α基因转移的ＮＩＨ３Ｔ３成纤维细胞（ＮＩＨ３Ｔ３－ＩＦＮ－α￣＋）就能显著抑制肝癌（皮下）体内生长，使荷瘤裸鼠存活期明显延长；（２）在腹腔埋植ＮＩＨ３Ｔ３－ＩＦＮ－α￣＋之后再静脉注射ＡＫ细胞、腹腔内注射ＩＬ－２，发现荷瘤裸鼠的皮下肿瘤结节生长抑制作用更明显；（３）将ＮＩＨ３Ｔ３－ＩＦＮ－α￣＋与ＩＬ－２／ＡＫ／阿霉素联合应用后，对肝癌裸鼠的治疗效果更佳。     

NK细胞的发育生物学

ＮＫ细胞是机体防御体系的第一道屏障。ＮＫ细胞的基本特性以及与另二类淋巴细胞关系的研究资料较少。本文重点阐述ＮＫ为核心的三类淋巴类细胞的共同前体细胞的发现，证明ＩＫＡＲＯＳ为淋巴类前体细胞的特有标志。继而证明ＮＫ是三类淋巴类细胞中更为接近Ｔ细胞的细胞。Ｔ／ＮＫ前体细胞存在于胎肝和胸腺中，可在不同诱导条件下定向分化为Ｔ或ＮＫ细胞。     

去甲基斑蝥素治疗卵巢癌的实验研究

目的 观察去甲基斑蝥素（ＮＣＴＤ）体内外对人卵巢癌细胞株３ＡＯ和ＡＯ的抑制作用。方法 利用细胞培养技术，以结晶紫染色测定；ＮＣＴＤ对３ＡＯ和ＡＯ细胞的体外杀伤作用，并在光镜和电镜下观察ＮＣＴＤ对其进行治疗，以观察ＮＣＴＤ体内对卵巢癌的抑癌效应。结果 体外实验表明ＮＣＴＤ对３ＡＯ和ＡＯ细胞有较强抑制作用，并呈浓度和时间依赖性。     

小剂量IL-2和IFNα与TNFα联合治疗恶性肿瘤的免疫学与临床研究

目的:研究小剂量细胞因子在恶性肿瘤免疫治疗中的意义。方法:全身应用小剂量重组干扰素(ＩＦＮα２ｂ)、白细胞介素－２(ＩＬ－２)和肿瘤坏死因子(α－ＴＮＦ)联合治疗多种恶性肿瘤６０例。应用ＡＰＡＡＰ法和免疫单向扩散法(ＲＩＤ)分析了治疗前后患者外周血中Ｔ、Ｂ淋巳细胞、ＮＫ细胞、免疫球蛋白(Ｉｇ)和补体Ｃ３活性。结果:治疗１个疗程后外周血中ＣＤ４＋的Ｔ淋巴细胞和ＣＤ２３＋的Ｂ淋巴细胞显著的高于治疗前水平(Ｐ＜０．０５);ＣＤ４＋/ＣＤ８＋比值也明显增高(Ｐ＜０．０１)。免疫球蛋白(ＩｇＭ)活性亦显著的高于治疗前水平(Ｐ＜０．０５)。ＫＰＳ积分较治疗前明显增高(Ｐ＜０．００１)。治疗后细胞因子加化疗组的ＫＰＳ积分平均增高２８．５分(Ｐ＜０．００１);ＣＲ ２６例(６５．０%);ＰＲ １１例(２７．５%),总有效率８２．５%。单纯细胞因子治疗组的ＫＰＳ积分平均增高２３分(Ｐ＜０．００１);ＰＲ １３例(６５．０%),总有效率６５．０%。结论:本方案可以显著的提高多种恶性肿瘤患者的细胞免疫和体液免疫。有可能提高肿瘤的化疗效果和患者的生存质量,延长生存期。     

乳腺癌患者手术前后免疫状况的测定

对３０例乳腺癌病人分别于术前、术后两周检测了静脉末梢血自然杀伤细胞（ＮＫ）活性和Ｔ淋巴细胞亚群状况，其中２０例于术后６周重复检测。结果显示，乳腺癌病人术前ＮＫ、ＯＫＴ４、ＯＫＴ４／ＯＫＴ８活性均降低，并随病情的进展呈减低趋势，在晚期病人中ＯＫＴ８增高尤其明显。术后２周与术前比较无显著性差异。术后６周ＮＫ、ＯＫＴ４、ＯＫＴ４／ＯＫＴ８均明显回升，而ＯＫＴ８显著降低。由此可见手术切除癌灶对病人的免疫恢     

特异性转移因子对荷瘤早期和晚期小鼠细胞免疫功能及生存期的影响

目的研究特异性转移因子（ＳＴＦ）对荷瘤小鼠的抗肿瘤作用。方法在小鼠腹腔内接种Ｓ１８０细胞同时腹腔注射５１８０－ＳＴＦ，测定天然杀伤（ＮＫ）和淋巴因子激活的杀伤（ＬＡＫ）活性、淋巴细胞转化（淋转）、ＬＡＫ特异杀伤活性及生存期。结果荷瘤小鼠ＮＫ和ＬＡＫ活性及淋转较正常小鼠明显低下（分别为１４．５５％±７１３％、１３．８２％±６．６４％、４４．１５±６．９４ＳＩ；２７．７４％±６．６７％、２９．３８％±９．１６％、６９．１４±７．６７ＳＩ），生存期明显缩短（分别为１７．５０±２．００ｄ和＞４５ｄ）。在荷瘤同时应用５１８０－ＳＴＦ，可部分阻止ＮＫ和ＬＡＫ活性及淋转的下降（分别为１８．６４％±７．１８％、２２．８４％±７．２２％和５３．０３±９．１５ＳＩ），延长生存期（２６．２５±４．４０ｄ），还可明显诱导ＬＡＫ特异活性。荷瘤３天后应用５１８０－ＳＴＦ，仅能部分诱导ＬＡＫ特异活性，不能延长生存期。结论ＳＴＦ可能有助于阻止癌前病变向癌转化，而对晚期肿瘤效果不佳。     

SITE SPECIFIC CANCER INDUCTION BY INTRA-ESOPHA-GEAL COTTON NODE RETENTION AND CARCINOGEN LADEN DRINKING WATER

ＳＩＴＥＳＰＥＣＩＦＩＣＣＡＮＣＥＲＩＮＤＵＣＴＩＯＮＢＹＩＮＴＲＡＥＳＯＰＨＡＧＥＡＬＣＯＴＴＯＮＮＯＤＥＲＥＴＥＮＴＩＯＮＡＮＤＣＡＲＣＩＮＯＧＥＮＬＡＤＥＮＤＲＩＮＫＩＮＧＷＡＴＥＲＬｕＪｉａｎｐｉｎｇ路建平ＨａｙａｓｈｉＫｅｉｋｉ林肇辉...     

A phase II study of the administration of recombinant interleukin 2 (rIL-2) plus lymphokine activated killer (LAK) cells in stage IV melanoma patients

From January 1987 to February 1988, 15 stage IV melanoma patients were treated with two courses of bolus injection of rIL-2 plus LAK cell infusions at the National Cancer Institute of Milan. The original treatment regimen included a first course of rIL-2 administration (400 micrograms/m2 bolus injection 3 times a day [TID] for 4 days) and a second course of rIL-2 administration (800 micrograms/m2 bolus injection TID for 7 days) separated by 4 consecutive daily leukaphereses. Autologous lymphokine activated killer (LAK) cells were reinfused into each patient on three occasions during the second period of rIL-2 administration. Due to the appearance of grade III-IV neurological, hepatic and cardiopulmonary toxicity, 7 patients discontinued dosing before the end of treatment, one patient desired to be withdrawn and one patient died from rapidly progressive disease, although complications of rIL-2 administration may have contributed to her death. Only 6 patients completed the schedule without evidence of major intolerance, even though the planned dose during the second course of rIL-2 was reduced to 400 micrograms/m2. The complete duration of treatment ranged from 11 to 19 days. The total dose of rIL-2 injected ranged from 12.6 to 30.4 mg. The number of infused LAK cells ranged from 15.5 x 10(9) to 60 x 10(9)/patient. Two of the 14 evaluable patients showed a minor anti-tumor response. In 5 patients new metastases in other sites were documented from 2 to 5 months after completion of dosing. No apparent association was found between progression of the disease (or the appearance of new metastases) and the total dose of rIL-2 injected, the number of LAK cells administered or the number of days of treatment. By December 1988, all patients had died of their disease in a period ranging from 3 to 14 months from the last injection of rIL-2. The lack of significant clinical responses in this study and the high toxicity of this treatment lead us to conclude that at least as far as melanoma patients are concerned, adoptive immunotherapy with rIL-2 plus LAK cells (as described here) is not a justifiable treatment option unless new evidence presents itself.     

晚期肺癌癌性胸水的免疫治疗

Ten patients with advanced lung cancer complicated by malignant pleural effusion were treated by intrapleural transfer of autologous LAK cells induced from lymphocytes of malignant effusions in the presence of rIL-2 and by administration of rIL-2 10 days before and after the transfer of LAK cells. The pleural effusions disappeared in 8 patients and significantly reduced in the other two. The number of tumor cells in the pleural effusion was obviously decreased while the number of lymphocytes was significantly increased. No changes were found in 4 responders during 4 months follow-up after treatment. No serious side effects were observed in all these 10 patients. The results indicated that transfer of LAK cells combined with rIL-2 in the treatment of patients with malignant pleural effusion due to advanced lung cancer is effective, safe and feasible.     

Recombinant interleukin-2 activates peripheral blood lymphocytes from children with acute leukemia to kill autologous leukemic cells

In order to determine if recombinant interleukin-2 (rIL-2) can induce lymphokine-activated killer (LAK) cells able to lyse autologous leukemic cells, we incubated peripheral blood (PB) mononuclear cells from children with acute leukemia with 50 U/ml rIL-2 for 5 days. These PB effector cells were then tested for their ability to lyse autologous leukemic cells in a 51CR release assay. The PB cells before incubation with rIL-2 showed little or no cytotoxicity for autologous blasts (range, 0 to 12%; mean, 2%). However, after incubation with rIL-2, PB cells from four of five children with acute lymphoblastic leukemia (ALL) at diagnosis or in relapse, and from six of eight children with ALL in remission were able to lyse autologous blasts. The percent lysis (range) was 0 to 69% (mean, 37%) for the former group, and 0 to 113% (mean, 43%) for the latter group. The PB cells from three patients (one in relapse and two in remission) failed to develop LAK activity after incubation with rIL-2. However, in each case cytotoxicity versus K562 was increased after incubation with rIL-2. Furthermore, in a Phase I study of rIL-2 for the treatment of refractory leukemia, a patient was treated with rIL-2 for 3 weeks (nine injections of 3 x 10(6) U/m2 each). Her fresh PB mononuclear cells developed a low level of cytotoxicity (11% lysis) against her autologous blasts during this time. The finding that rIL-2 in vitro and in vivo induces LAK cells with cytotoxicity against autologous leukemic cells provides the rationale for the clinical trial of this agent in the treatment of children with ALL.     

Adoptive immunotherapy for recurrent glioblastoma multiforme using lymphokine activated killer cells and recombinant interleukin-2

Thirteen patients with recurrent glioblastoma were treated with adoptively transferred autologous lymphokine activated killer (LAK) cells and recombinant interleukin-2 (rIL-2). Patients' blood mononuclear cells (MNC) obtained by leukapheresis were cultured at 2.5 million MNC per ml for 3 to 5 days in media containing 1000 U rIL-2/ml. After incubation, the nonadherent MNC from all cultures (0.5-5 X 10(9] were combined and concentrated for infusion in 5 to 10 ml saline containing 10(6) U rIL-2. Nine patients received one injection of LAK cells and rIL-2 into the brain tissue immediately surrounding the tumor cavity during craniotomy for subtotal tumor removal (Group 1). On each of the 3 days after surgery, patients received boosters of 10(6) U rIL-2 delivered into the tumor cavity through a skin flap or via an Ommaya reservoir. Approximately 1 to 2 weeks after this series of injections, these patients were treated with a second cycle of LAK cells and rIL-2 injected into the tumor cavity using the reservoir. Four patients received both adoptive immunotherapy cycles by intracavitary injection (Group 2). In this relatively small patient pool, neither age, sex, Karnofsky score, treatment history, nor anticonvulsant and steroid dosage appeared to influence a patient's ability to make LAK cells. The therapy, itself, was well-tolerated by all patients although they all displayed symptoms of aseptic meningitis and increased intracranial pressure, i.e., headache, fever, malaise on the days of LAK cell and/or rIL-2 infusion. The therapy did not appear to have a significant impact on patient survival (mean, 30 weeks) especially for those patients with a high postsurgical tumor burden. As the therapy is safe, the authors believe its efficacy can best be tested in patients with a newly diagnosed or recurrent glioblastoma which lies in an area where a near-total resection is possible.     

TheIn vitro potentiation of LAK cell cytotoxicity in cancer and aids patients induced by F3—A fractionated extract of astragalus membranaceus

Thein vitro induction of LAK cell activity was studied in cancer and AIDS patients. F3, an immuno-regulatory component of Astragalus membranaceus was shown capable of potentiating LAK cell activity induced by rIL-2. The LAK cells killing activity against Hs294T melanoma cell line induced by 50 U/ml rIL-2 in the presence of F3 (55 μmg/ml) reached 64%, which was comparable to that (60%) induced by 500 U/ml of rIL-2 alone. With F3 and rIL-2, the effector to target ratio could be reduced to one-half in order to obtain an equivalent level of cytotoxicity induced by rIL-2 alone. In some patients whose peripheral blood lymphocytes were relatively inert of rIL-2, F3 could make them responsive to rIL-2 induction. These results imply that F3 may be useful to potentiate LAK cell activity, reduce the dosage of rIL-2 and thus minimize the later’s toxic side effects when usedin vivo.     

Phase I trial of intraperitoneal recombinant interleukin-2/lymphokine-activated killer cells in patients with ovarian cancer

Ten patients with ovarian cancer refractory to conventional therapy were treated with intraperitoneal (i.p.) recombinant interleukin-2 (rIL-2) and lymphokine-activated killer cells (LAK). The 28-day protocol consisted of 6 priming i.p. rIL-2 infusions on days 0, 4, 6, 8, 10, and 12. Leukapheresis was performed for mononuclear cell collection on days 15, 16, 17, and 18 and lymphokine-activated killer cells were given i.p. with the rIL-2 on days 19 and 21. Three additional i.p. rIL-2 infusions were given on days 23, 25, and 27. Three dose levels of rIL-2 were tested: 5 X 10(5), 2 X 10(6), and 8 X 10(6) units/m2 body surface area. The dose-limiting toxicity was abdominal pain secondary to ascites accumulation with significant weight gain. Other toxic effects included decreased performance status, fever, nausea and vomiting, diarrhea, and anemia. Peripheral lymphocytosis and eosinophilia were seen at all dose levels. The maximum tolerated dose is 8 X 10(6) units/m2/dose. Peripheral and peritoneal IL-2 levels were measured with a bioassay using an IL-2-dependent cell line. At the highest dose level, serum IL-2 was greater than 10 units/ml for 18 h. After the first infusion, a 2-log dilution of the i.p. IL-2 was measured in the serum. In the postleukapheresis i.p. IL-2-dosing period less IL-2 was detected in the serum than in the earlier i.p. IL-2-priming period. The induction and persistence of LAK activity were studied. Peritoneal LAK activity was detected as early as 4 days after the first i.p. infusion, by day 11 in all evaluable patients, and persisted for the 6-day interval between priming IL-2 and LAK/IL-2 infusion. Peritoneal lytic activity persisted until day 28 in 5 tested patients. These peritoneal cells retained lytic activity 48 h in culture medium without rIL-2 present. Peritoneal LAK activity correlated with the percentage of mononuclear cells and the percentage of CD56-positive mononuclear cells in the peritoneum. The yield of peripheral lymphocytes after the six i.p. priming doses of rIL-2 correlated with the dose level of rIL-2 infused. Peripheral blood LAK activity showed a minimal, however progressive, increase during the treatment protocol. LAK activity could be enhanced if rIL-2 was present during the 4-h assay. These studies indicate that i.p. rIL-2 infusion induced durable regional LAK activity and primes peripheral blood cells for LAK activity if exposed briefly to additional IL-2.     

Inducible lymphokine-activated killer (LAK) cell activity in the peripheral blood of patients with relapsed/refractory non-Hodgkin's lymphoma (NHL)

Therapy with recombinant interleukin-2 (rIL-2) induces clinical response in a significant number of patients with refractory malignant disease. Very few patients with non-Hodgkin's lymphoma (NHL) have been treated with rIL-2. The present study sought to determine if peripheral blood mononuclear cells (PBM) from patients with relapsed/refractory non-Hodgkin's lymphoma could be induced in vitro to generate LAK cell activity. PBM from 28 patients with relapsed/refractory NHL were incubated for 7 days in rIL-2 to determine their ability to lyse the LAK cell sensitive Daudi cell line. The PBM from all patients were able to generate LAK activity after in vitro incubation in rIL-2. Approximately one third of the patients' PBM samples generated less activity than activity generated in the PBM sample from normal control donors. However, two-thirds of patient samples were able to generate activity equal to or greater than that of the controls. The degree of LAK activity generated by the patients' PBM did not correlate either with histologic subtype or amount of prior chemotherapy. The amount of LAK activity an individual generated (control or patient) tended to remain stable over time.     

Effect of anti-CD3 antibody on the generation of interleukin-2-activated lymphocytes from tumor tissues of gastrointestinal cancer.

BACKGROUND. The efficiency of anti-CD3 antibody (OKT3) for adoptive immunotherapy using lymphokine-activated killer (LAK) cells generated from tumor-infiltrating lymphocytes (TIL), regional lymph node lymphocytes (RLNL), and peripheral blood lymphocytes (PBL) was investigated. METHODS. TIL, RLNL, and PBL derived from 39 patients with gastrointestinal cancers (16 gastric cancers, 17 colorectal cancers, and 6 esophageal cancers) were cultured for 4 weeks with 200 U/ml of recombinant interleukin-2. To one group, solid-phase 10 micrograms/ml OKT3 was added during the initial culture period (day 2 or 4). Cytotoxicity against K562 cells (NK-like activity) and Daudi cells (LAK activity) and the phenotypes of effector cells generated after culturing for 2-3 weeks were studied. RESULTS. Proliferative responses were significantly increased by OKT3 in each type of effector cell (P less than 0.01); in particular, TIL expanded more by OKT3 than PBL and RLNL (P less than 0.01). The population of CD8+ CD11b- cytotoxic T-cells in OKT3-stimulated groups was significantly larger than that in unstimulated groups (P less than 0.01), whereas no differences were observed with CD4+ cells (helper/inducer T-cells) and CD8+ CD11b+ cells (suppressor T-cells). OKT3 enhanced the NK-like activity of TIL and PBL but did not affect their LAK activity. OKT3 suppressed the NK and LAK activity of RLNL. CONCLUSIONS. OKT3 stimulation did not significantly enhance the LAK activity, but the authors propose that OKT3 could be an effective addition to adoptive immunotherapy using TIL due to an increased proliferation and generation of a large cytotoxic T-cell population.     

rIL－2/LAK细胞治疗晚期恶性肿瘤

 胎儿来源的LAK细胞/rIL-2对5例失去手术、化疗、放疗机会的晚期恶性肿瘤病人进行了过继免疫治疗。结果都取得了-定的疗效,黑色素瘤病人肺转移结节基本消失,局部瘤细胞全部坏死,组织细胞正常;淋巴瘤病人转移灶消退59%;小肠平滑肌肉瘤病人在没进行此疗法之前2年半复发、手术三次。第三次手术后用此疗法治疗-个疗程,三年后第-次复发。为晚期恶性肿瘤病人延长了生命,提高了生存质量争得了再次治疗的机会。     

In vivo and in vitro activation of natural killer cells in advanced cancer patients undergoing combined recombinant interleukin-2 and LAK cell therapy

Patients with advanced metastatic cancer were given combined autologous lymphokine activated killer (LAK) cell and recombinant interleukin-2 (rIL-2) therapy on a National Cancer Institute extramural phase II trial. Systemic administration of rIL-2 resulted in pronounced lymphocytopenia. Within two days after completion of in vivo rIL-2 therapy, there was a dramatic increase in absolute numbers of circulating lymphocytes, and cytotoxic activity against tumor cell targets was mediated by peripheral blood lymphocytes, indicating in vivo generation of LAK activity. Patients were leukapheresed and cells cultured for three to four days in rIL-2. rIL-2 cultured cells from all patients demonstrated cytotoxic activity. In order to characterize the effector cell, T cells and natural killer (NK) cells were isolated to greater than 95% purity by flow cytometry. Cytotoxic activity was mediated by rIL-2--activated NK cells, whereas T cells demonstrated no substantial activity. The circulating in vivo cytotoxic effectors detected after in vivo rIL-2 therapy were also shown to be rIL-2--activated NK cells. Results from these studies demonstrate that all patients were capable of generating a cytotoxic response, and that the cytotoxic effector cells were rIL-2--activated NK cells, identified by the phenotype CD3--, Leu 19+.