LPAK细胞／IL—2治疗肿瘤的初步临床观察

用ＬＰＡＫ（ＬｙｍｐｈｏｋｉｎｅａｎｄＰＨＡＡｃｔｉｖｉｔｅｄＫｉｌｌｅｒ）细胞／ＩＬ－２过继免疫疗法治疗１７例晚期肿瘤，多数病人治疗后有不同程度的症状改善，如肺癌病人刺激性咳嗽、胸疼减轻，肝癌病人肝区疼痛明显减轻或完全消失。在１２例可评估病例中，５例肝癌（包括转移性肝癌１例）有３例肿瘤分别缩小３５．７、３９．５和４７．８％，其中１例在治疗过程中门静脉一２．７×１．４ｃｍ的癌栓完全消失。２例肺癌和２例转移性肺癌治疗后肿瘤无明显变化，１例胃癌癌性腹水治疗后腹水大部消失，腹围由治疗前的８２ｃｍ缩小至７５ｃｍ，１例皮肤恶性淋巴瘤治疗后皮损消退５０％以上。我们观察了其中６例肿瘤病人在治疗前后的淋巴细胞转化，淋巴细胞对ＩＬ－２、淋巴细胞对ＩＬ－２和ＰＨＡ联合刺激的反应性以及ＮＫ活性，发现治疗后病人的前３项指标均有显著提高（Ｐ＜０．０１）ＮＫ活性也有增高趋势。ＬＰＡＫ细胞的主要副作用（实际包含ＩＬ－２的副作用）有：短暂发热（３９℃左右），寒战，腹痛、纳差，体液潴留等，不经处理或对症处理后上述症状可消失。     

原发性肝癌肝动脉化疗栓塞合并LAK/IL-2灌注治疗的临床Ⅲ期观察

评价肝动脉化疗栓塞合并ＬＡＫ／ＩＬ┐２灌注治疗原发性肝癌的初步疗效。方法不能切除的肝癌患者经肝动脉化疗栓塞后灌注自体ＬＡＫ细胞及ＩＬ┐２，并与单独行肝动脉化疗栓塞的不能切除肝癌患者比较其肿瘤大小，生活质量的变化及毒副反应。结果经肝动脉化疗栓塞合并ＬＡＫ／ＩＬ┐２灌注治疗的２２例肝癌有效率（ＣＲ＋ＰＲ）为１３．６％，包括１例完全缓解和２例部分缓解，而单独化疗栓塞对照组１７例中仅１例显示部分缓解（有效率５．９％）。化疗栓塞＋ＬＡＫ／ＩＬ┐２治疗组与单独化疗栓塞对照组中ＭＲ、ＳＤ、ＰＤ分别为５、１２、１例和１、１１、４例。治疗组中大多数病人生活质量改善或稳定，而对照组中生活质量则无提高。两组病人的毒副反应均较轻而短暂。结论经肝动脉化疗栓塞合并ＬＡＫ／ＩＬ┐２灌注治疗原发性肝癌疗效较优于单独肝动脉化疗栓塞治疗，原发性肝癌更有效的综合治疗方案有待进一步探索     

人天然白介素－2治疗恶性肿瘤的Ⅱ期临床观察

目的评价人天然白介素－２（ＩＬ－２）治疗恶性肿瘤的效果。方法采用ＩＬ－２加淋巴因子激活的杀伤细胞（ＬＡＫ细胞）全身输注治疗１４例实体瘤（恶性黑素瘤４例，肾癌２例，原发性肝癌和肠癌各３例，恶性淋巴瘤和恶性腮腺混合瘤各１例）和胸腹腔内注入治疗１０例癌性胸腹水（胃癌４例，卵巢癌２例，肠癌、乳腺癌、胰腺癌和肺癌各１例）。结果实体瘤缓解率为２１．４％（３／１４），胸腹水缓解率为７０％（７／１０）。不良反应有发热（＜３９℃）３例；恶心１例；肾功能障碍１例，停药后恢复正常。结论人天然ＩＬ－２具有一定的抗肿瘤作用。     

LAK细胞和IL－2联合化疗治疗晚期肺癌的近期疗效观察

ＬＡＫ细胞和ＩＬ－２是目前常用的肿瘤生物制剂。自１９９２年以来，对１８例晚期肺癌，男１５例，女３例，年龄２９～６４岁，进行ＬＡＫ细胞和重组ＩＬ－２联合化疗治疗。选择胎脾、胸腺的淋巴细胞做前体细胞，体外用重组ＩＬ－２诱导制备ＬＡＫ细胞，每输３次ＬＡＫ细胞为一疗程、每次输入细胞数０．５～１．０×１０９。化疗采用环磷酰胺，长春新碱，阿霉素为主的方案。治疗结果：完全缓解（ＣＲ）５例，部分缓解（ＰＲ）７例，无效（ＮＲ）３例，病情平稳３例，有效率ＣＲ＋ＰＲ达６６％。采用本疗法后病人精神及饮食好转，能有效缓解胸痛、减轻病人痛苦。提示ＬＡＫ细胞和重组ＩＬ－２联合化疗对晚期肺癌是一种可行的有效治疗，但在临床应用中要注意毒性反应。     

经肝动脉门静脉输注自体LAK细胞／IL－2及化疗药物治疗7例中晚期肝癌的临床观察

我院１９９３年４月至６月经肝动脉及门静脉安置的植入式微泵，输注自体ＬＡＫ细胞／ＩＬ－２及化疗药物治疗７例中晚期肝癌（其中２倒置泵，同时进行了肿瘤姑息性切除），近期疗效ＣＲ２例，ＰＲ３例，ｍＲ１例，ＰＤ１例，总有效率达８５．７％，且毒性反应轻微。结果提示：对于中晚期肝癌，经肝动脉门静脉输注自体ＬＡＫ细胞／ＩＬ－２及化疗药物，是一种极有前途的治疗措施，可望明显提高肝癌疗效。     

白介素－2的Ⅱ期临床试验报告

共１５９例各种类型的恶性肿瘤病人进入了Ⅱ期临床试验。２５例病人接受了白介素－２（ＩＬ－２）单独全身治疗（静脉或皮下注射）；６０例病人接受了ＩＬ－２＋ＬＡＫ细胞全身性治疗；４１例癌性胸水病人接受了ＩＬ－２胸腔内灌注；６例癌性腹水病人接受了ＩＬ－２腹腔灌注；２７例病人接受了ＩＬ－２瘤内注射。单用ＩＬ－２全身治疗的病人没有病例获得临床缓解，２３晚期肾癌接受了ＩＬ－２＋ＬＡＫ细胞治疗，５例获得部分缓解，有效率２２％：癌件胸水和腹水的有效率分别为６８％和６７％：ＩＬ－２瘤内注射的有效率为３０％。Ｔ４／Ｔ８比值、ＮＫ细胞活性和ＬＡＫ细胞活性在全身性ＩＬ－２治疗后均有显著升高，并有统计学意义。病人接受本试验所用的ＩＬ－剂量后的毒副反应主要为发热、畏寒或寒战、疲乏，全组病人均无发生严重低血压、液体潴留等毛细血管渗漏现象。     

不能切除的肝癌术中治疗模式的探讨（附200例随访结果）

自１９６４年４月～１９９３年７月对２００例不能切除的肝癌术中应用多种模式治疗。患者均经手术及病理证实。按治疗方法分为两组。Ⅰ组：１１５例单纯作肝动脉插管化疗（ＨＡＩ），其１、２年生存率分别为１０．４％和１．７％，仅１例生存６５个月；Ⅱ组：８５例术中作肝动脉栓塞化疗（ＨＡＣＥ）和加用肝动脉结扎（ＨＡＬ）或伴有微波刀肿瘤固化（ＭＩＣ）、或伴用纯酒精肿瘤内注射（ＥＩＴ）的多模式治疗，其中１、２、３和５年生存率分别为３４．１％、２１．２％、１２．０％和６．７％，５例仍生存４１～６３个月，６例肿瘤缩小后获二期切除。比较两组的治疗结果，对不能切除的肝癌术中采用多模式的治疗，是延长其生存率的有效方法。结合文献复习，对多模式的治疗原理进行了讨论。     

枸杞多糖联合LAK／IL－2疗法对75例晚期肿瘤的疗效观察

用枸杞多糖（ＬｙｃｉｕｍＢａｒｂａｒｕｍＰｏｌｙｓａｃｃｈａｒｉｄｅｓ，ＬＢＰ）联合ＬＡＫ／ＩＬ－２疗法临床试验治疗７９例晚期肿瘤患者，其中７５例可评估病人资料分析提示，ＬＢＰ联合ＬＡＫ／ＩＬ－２疗法组疗效（４０．９％）显著优于ＬＡＫ／ＩＬ－２疗法组（１６．１％）。两种治疗方案对恶性黑色素瘤、肾癌、直结肠癌、肺癌、恶性胸水和鼻咽癌有一定的疗效。ＬＢＰ联合ＬＡＫ／ＩＬ－２治疗组的缓解持续时间显著长于ＬＡＫ／ＩＬ－２治疗组。ＬＢＰ联合ＬＡＫ／ＩＬ－２治疗组治疗前后外周血淋巴细胞（ＰＢＬ）的ＮＫ、ＬＡＫ活性增高程度均显著大于ＬＡＫ／ＩＬ－２治疗组，说明ＬＢＰ能够提高ＬＡＫ／ＩＬ－２疗法对晚期肿瘤的治疗效果。     

肝癌切除术后栓塞化疗与LAK细胞／IL－2的作用

目的探讨肝癌切除术后进行栓塞化疗或（和）淋巴因子激活的杀伤细胞（ＬＡＫ）／白介素－２（ＩＬ－２）的远期疗效。方法回顾性分析肝癌切除术后３年以上患者１０４例，比较单纯切除（Ａ组）、切除加栓塞化疗（Ｂ组）和切除加栓塞化疗加ＬＡＫ细胞／ＩＬ－２（Ｃ组）的预后情况。结果３年生存率在Ａ、Ｂ、Ｃ组分别为３０％、５４．３％、５７．１％。根治性切除病例，Ａ、Ｂ、Ｃ组的３年复发率分别为７６．７％、４６．７％、４１．７％。单纯手术组的生存率显著低于其他两组，而根治性切除后的复发率则显著比另两组为高。结论肝癌切除联合栓塞化疗或（和）ＬＡＫ细胞／ＩＬ－２治疗有助于改善肝癌患者的预后。     

异环磷酰胺治疗晚期非小细胞肺癌的疗效观察

「目的」观察异环磷酰胺（ＩＦＯ）为主的方案治疗晚期非小细胞肺癌的疗效和毒副反应２。「方法」采用ＩＡＰ或ＩＥＰ方案治疗晚期非小细胞肺癌３８例，ＩＦＯ１．４ｇ／ｍ＾２ｄ１￣５；ＤＤＰ２５ｍｇ／ｍ＾２ｄ１￣３；ＡＤＭ３０ｍｇ／ｍ＾２ｄ１和Ｖｐ１６６０ｍｇ／ｍ＾２ｄ１￣３。「结果」鳞癌有效率３５．７％（５／１４）；腺癌有效率为４５．０％（９／２０）；总有效率３９．５％（１５／３８）。主要毒副反应为骨髓抑制     

Biological, histological, and clinical impact of preoperative IL-2 administration in radically operable gastric cancer patients

BACKGROUND AND OBJECTIVES: Surgery induces lymphocytopenia and this decrease of host defenses, related to interleukin-2 (IL-2) endogenous imbalance during postoperative period could promote the proliferation of possible micrometastases and the implantation of surgically disseminated tumor cells. Moreover, tumor infiltrating lymphocytes (TILs), activated by endogenous IL-2 release, are linked to prognosis in cancer patients. The aim of this randomized study is to assess the biological (peripheral blood cells count, related to the grade of immunosuppression), histological (TILs), and clinical (overall and disease-free survival) impact of preoperative low doses administration of IL-2 in patients with radically operable gastric cancer. METHODS: This prospective study enrolled 69 consecutive patients with histologically proven gastric adenocarcinoma who underwent radical surgery from October 1999 to December 2002 (M/F 39/30; mean age 66; range 42-82) who underwent radical surgery from October 1999 to December 2000. Patients were randomized to be treated with surgery alone as controls (35 patients) or surgery plus preoperative treatment with recombinant human IL-2 (34 patients). We considered the total lymphocyte count and lymphocyte subset (CD4, CD4/CD8) during the preoperative period, before IL-2 administration, and on the 14th and 50th day, peritumoral stromal (fibrosis) reaction, neutrophils, lymphocytes, and eosinophils infiltration in tumor histology, and morbidity disease free and overall survival were evaluated. RESULTS: Two groups were well matched for type of surgery and extent of disease. All the patients underwent radical surgery plus D2 lymphadenectomy. At baseline, there were no significant differences in total lymphocyte and lymphocyte subsets between groups. The control group showed a significant decrease of total lymphocytes, CD4 cells, and CD4/CD8 ratio at the 14th postoperative day relative to the baseline value. In the control group 65% of patients had a decrease of CD4 under 500 cells/mmc. Instead it has been observed in IL-2 group a significant increase over the control group values of total lymphocytes and CD4 cells (14th total lymphocytes and CD4: IL-2 vs. control P < 0.05). Moreover in this group only 15% patients had CD4 under 500 cells/mmc. This difference, in CD4 count, is significant even at the 50th postoperative day (P = 0.006). IL-2 group showed lower postoperative complications (2/34 vs. 11/35; P < 0.05), and higher lymphocyte/eosinophil infiltration into the tumor (P < 0.0002). Median follow-up was 26 months (range 10-48) and median overall and disease-free survivals were longer, even if not significantly, in the IL-2 group than in the control arm (P = 0.07 and P = 0.06 respectively). CONCLUSIONS: This randomized study would suggest that a preoperative immunotherapy with IL-2 is a well-tolerated treatment able to prevent surgery-induced lymphocytopenia. IL-2 seems to neutralize the immunosuppression induced by operation and so to stimulate the host reaction against tumor tissue (lymphocytes/eosinophils infiltration). Furthermore IL-2 seems to have an impact on clinical course reducing morbidity of surgery and ameliorating overall and disease-free survival.     

肺瘤平膏及其拆方对DC刺激LPAK抗肿瘤活性的影响

目的 比较不同中药含药血清对树突状细胞(dendritic cell,DC)刺激LPAK抗肿瘤活性,为指导临床用药提供初步的实验依据.方法从健康人外周血中分离获得PBMC,采用多种细胞因子诱导,获取DC及LPAK,采用中性红摄入比色法检测肺瘤平膏及其拆方含药血清干预DC LPAK细胞杀伤肿瘤细胞活性的不同.结果 LPAK∶Tumor(L∶T)为10∶1或5∶1组时,各中药组DC诱导的LPAK细胞,杀伤活性明显高于对照组(P＜0.05).组内杀伤活性比较,即L∶T(10∶1)与L∶T(5∶1)比较,解毒组、空白DC对照组、T+ LPAK组,P＜0.05,余各组比较,P＞0.05;肺瘤平膏组、益气组、活血组在L∶T为5∶1与10∶1时,诱导LPAK的杀伤活性基本相同.结论 肺瘤平膏、活血药、益气药可不同程度增强DCLPAK杀伤肿瘤细胞的作用,而解毒药则对其有抑制作用.     

肿瘤负荷对CIK/IL-2临床疗效的影响

目的:自从1993年Schmidt Wolf建立细胞因子诱导的杀伤细胞(CIK)制备方法以来,CIK/IL-2在净化骨髓和治疗血液病方面取得较多经验,但缺乏CIK/IL-2治疗实体瘤的临床资料.本文总结应用CIK/IL-2治疗41例实体瘤的临床经验,分析肿瘤负荷对该疗法临床疗效的影响.方法:选择无临床病灶的高危复发患者和存在明显临床病灶的晚期实体瘤患者,分离外周血单个核细胞,制备CIK.CIK联合IL-2静脉输注患者.结果:高危复发组中,姑息性治疗9例,根治性治疗14例,中位随访时间13个月.接受CIK/IL-2治疗后姑息性治疗者复发6例,根治性治疗者复发3例,复发率分别为66.7%和21.4%(P＜0.05).此外,CIK/IL-2治疗常规治疗失败的晚期实体瘤患者18例,4例稳定.结论:CIK/IL-2的临床疗效受肿瘤负荷制约,接受CIK/IL-2辅助治疗前应联合其它治疗方法尽量降低肿瘤负荷.     

白介素6在雌激素促进小鼠肺腺癌进展中的作用及其机制

目的 探讨白介素6（interleukin 6, IL-6）在雌激素（estrogen, E2）促进小鼠肺腺癌进展中的作用及其可能的机制。方法 采用乌拉坦诱导雌性昆明小鼠建立肺腺癌模型,进行如下分组处理（每组9只）：空白对照组、E2组、雌激素抑制剂（E2 inhibitor, E2I）组、E2+E2I组、IL-6组、E2+IL-6组。小鼠饲养16周后处死检测小鼠体重变化、成瘤情况、肿瘤分级、肺脏器指数等指标,酶联免疫法（ELISA）检测对照组血清标本中E2/IL-6的含量,实时荧光定量PCR（Real-Time PCR, RT-PCR）检测对照组肿瘤标本中ERβ/IL-6的mRNA表达水平,采用免疫印迹法检测各组ERβ、Akt、MAPK,p-ERβ、p-Akt、p-MAPK表达水平。结果 肺部出现结节的昆明小鼠占小鼠总数的比例（即称为成瘤率）为87.04%（47/54）,小鼠肺部结节做病理切片后证实为肺腺癌的小鼠数目占小鼠总数的比例（即成癌率）为70.37%（38/54）,肺结节数、肿瘤指数、肺脏器指数等统计指标在E2组显著高于E2+E2I组、E2I组、E2+IL-6组、IL-6组及空白对照组（P均<0.05）;对照组小鼠血清中E2与IL-6呈高度相关性（P均<0.05）,各组小鼠肺癌组织中Akt、MAPK、ERβ、p-AKt、p-MAPK、p-ERβ的表达水平及ERβ的mRNA表达水平与肿瘤统计指标表达的趋势相一致（P均<0.05）。结论 在E2促进小鼠肺腺癌进展过程中IL-6具有下调ERβ信号通路作用,提示IL-6可能抑制E2促进肺腺癌进展。     

IL-12- and IL-2-induced tumor regression in a new murine model of oral squamous-cell carcinoma is promoted by expression of the CD80 co-stimulatory molecule and interferon-gamma

Therapy with IL-12 or IL-2 induces tumor regression in only a few patients with head-and-neck squamous cell carcinoma (SCC), and the factors promoting responsiveness have not been well defined. In this study, we examined whether combined IL-12 and IL-2 therapy can induce tumor regression in a new murine model of oral SCC and determined if the anti-tumor response is promoted by expression of the immune co-stimulatory molecule CD80 and cytokine IFN-gamma. In CD80-positive or -negative subclones of a BALB/c oral SCC line in syngeneic mice, we showed that systemic rIL-12 alone was comparable in effectiveness to combined therapy with IL-12 and peri-tumoral rIL-2, inducing complete regression of the CD80(+) line B7E11-4scid. However, therapy with these cytokines had no effect on growth of the CD80(-) subclone B7E3-4scid and did not induce complete regression of the CD80(+) subclone B7E11-4scid in congenic BALB/c IFN-gamma knockout mice, indicating that expression of the CD80 co-stimulatory molecule and IFN-gamma contributes to tumor regression. In cytokine-treated mice that rejected the CD80(+) SCC line, an increase in infiltrating CD4(+) lymphocytes and apoptotic bodies within the tumor specimens was observed, and resistance to rechallenge with the same tumor was detected in 50% of recipients, consistent with an immune response. Our results provide evidence that regression of oral head-and-neck SCC may be induced by therapy with systemic IL-12 and that expression of the CD80 co-stimulatory molecule by SCC and IFN-gamma by the host promote IL-12 induced regression of SCC.     

Clinical efficacy of adoptive immunotherapy by IL-4 activated tumor-infiltrating lymphocytes in patients with advanced cancer

Background Adoptive immunotherapy using tumor-infiltrating lymphocytes (TILs) has been used to treat malignant melanoma and renal cell carcinoma, although the clinical efficacy of this method has not yet been proved. To improve clinical efficacy, it is important to induce sufficient amounts of tumor-infiltrating lymphocytes to fight the tumor. In this study, we investigated the clinical efficacy of adoptive immunotherapy using interleukin (IL)-2 activated tumor-infiltrating lymphocytes combined with IL-4. Methods Tumor-infiltrating lymphocytes from patients with non-malignant melanoma and non-renal cell carcinoma were cultured with IL-2 monotherapy (n=10) and combination therapy with IL-4+IL-2 (n=20). Comparing the 2 groups, we investigated the clinical benefits of adoptive immunotherapy, considering safety, clinical efficacy, survival periods, and quality of life. Results By using the IL-4+IL-2 combination, we successfully transferred 6.5 times more activated tumor-infiltrating lymphocytes, as compared to cell counts using IL-2 monotherapy. The response rate achieved was 58.8% (2 complete and 8 partial responses) without any side effects in the IL-4+IL-2 group. Furthermore, the IL-4+IL-2 group had longer survival periods and improved quality of life, compared to the IL-2 monotherapy group. Conclusion The IL-4+IL-2 combination improved the clinical efficacy of adoptive immunotherapy, and improved quality of life in patients with non-malignant melanoma and non-renal cell carcinoma.