Effect of a streptococcal preparation (OK432) on natural killer activity of tumour-associated lymphoid cells in human ovarian carcinoma and on lysis of fresh ovarian tumour cells

The streptococcal preparation OK432 was studied for its effects on natural killer (NK) activity of peripheral blood lymphocytes (PBL) from normal donors and from ovarian cancer patients, and of tumour-associated lymphocytes (TAL) from peritoneal effusions. OK432 augmented NK activity against the susceptible K562 line and induced killing of the relatively resistant Raji line. Freshly isolated ovarian carcinoma cells were relatively resistant to killing by unstimulated PBL and TAL. OK432 induced significant, though low, levels of cytotoxicity against 51Cr-labelled ovarian carcinoma cells. Augmentation of killing of fresh tumour cells by OK432 was best observed in a 20 h assay and both autologous and allogeneic targets were lysed. PBL were separated on discontinuous Percoll gradients. Unstimulated and OK432-boosted activity were enriched in the lower density fractions where large granular lymphocytes (LGL) and activity against K562 were found. Thus, OK432 augments NK activity of PBL and TAL in human ovarian carcinomas and induces low, but significant, levels of killing of fresh tumour cells. Effector cells involved in killing of fresh ovarian tumours copurify with LGL on discontinuous gradients of Percoll.     

B型超声在肝癌手术中的应用

本文报告B超在11例肝癌手术中的应用。共发现18个癌灶。最大直径15cm,最小直径0,2cm。大于3cm的9个癌灶已被术前B超,CT和术中B超检出,其中4例经探查大体判断无法切除,但在术中B超引导下切除成功。小于2cm的9个癌灶,术前B超、CT无一发现,其中4个虽经手术探查亦未能看到或摸到,经由术中B超检出。本文详细描述了肝癌与脉管间的关系和癌灶的声像图特征,同时还就术中B超对肝癌诊断及手术应用价值进行了讨论。     

CIK细胞体内外抗肝癌细胞作用

目的：研究肝癌患者CIK（cytokine-induced killer) 细胞的体外杀伤自体肝癌原代细胞的细胞毒活性以及正常人CIK细胞在裸鼠体内的抗肿瘤作用。方法：分别分离获得肝癌患者和下人的外周血单个核细胞（PBMC）,加入细胞因子,体外诱导成CIK细胞,用流式细胞仪对细胞作动态表型分析,并与正常人的CIK细胞作对比。用^51Cr释放法,测定肝癌患者的CIK细胞体外杀伤自体肿瘤细胞的细胞毒性活性。在Balb/c裸鼠皮下接种肝癌细胞BEL－7402,观察CIK细胞对荷瘤鼠的抑瘤作用,并与LAK、PBMC细胞相对比。结果：肝癌患者的CIK细胞体外增殖力强,至培养28天时达到最大增值倍数300多,表型分析结果表明,CD^3 CD56^ 双阳性细胞得到了大量的扩增,其含量由原来的0．23％上升到第21天的17．8％。体外实验表明,肝癌患者的CIK细胞杀伤自体原代肝癌细胞的细胞毒性活性明显高于自体的PBMC细胞。裸鼠体内实验表明,肝癌患者的CIK细胞能够显著抑制肿瘤的生长,其抑瘤率可达84．7％,高于LAK细胞的52．8％及PBMC的37．1％（P＜0．05和P＜0．01）。结论：CIK细胞具有较强的体内外抗肝癌细胞活性,有可能应用于临床上肝癌的过继性免疫治疗。     

Effects of Recombinant Interferon-Gamma and Interleukin-2 on the Generation of Lymphokine-Activated Killer Cells in Vitro

In order to determine if recombinant interferon-γ (rIFN-γ) can augment the effect of recombinant interleukin-2 (rIL-2) in generating lymphokine-activated killer (LAK) cells, we have incubated normal peripheral blood mononuclear cells (PBMC) with these lymphokines for 3 days and then tested their LAK and natural killer (NK) cell activity. We have found that LAK activity in PBMC from 13 out of 13 normal donors was increased by the combined lymphokines above that due to either lymphokine alone, provided that rIL-2 was present at suboptimal concentration: Optimal levels of rIFN-γ (100 U/ml) were able to enhance the LAK-inducing activity of suboptimal levels (5 U/ml) but not optimal levels (100 U/ml) of rIL-2. NK activity showed a similar response to these concentrations of lymphokines. Activation of LAK/NK cells was accompanied by increases in the percentages of Leu 19⁺ (CD56) cells and TAC⁺ (IL-2-receptor) cells, and in the intensity of TAC antigen expression. These results indicate that combination rIFN-γ and rIL-2 may be more effective in generating LAK/NK cells than rIL-2 alone, particularly with suboptimal concentrations of rIL-2 such as occur during continuous infusion therapy with this agent.     

Phenotypical analysis of effector cells on nonspecific cancer cell therapy

In the present study, we examined the contributions of lymphocyte subpopulations in lymphokine activated killer (LAK) activity. LAK cells prepared from peripheral blood mononuclear cells (PBMC) of healthy donors showed highly cytotoxic activities against target tumor cells. When CD16 and CD56 positive cells in LAK cells were depleted by magnetic cell sorting, their cytotoxic activities were dramatically decreased. In contrast, little change was observed by the depletion of CD3 positive cells, suggesting that CD16 and/or CD56 positive populations, but not CD3 positive populations, including natural killer (NK) cells are the major cell types involved in LAK activity. Indeed, NK-enriched LAK cells prepared by culturing PBMC with IL-2 and OK-432 showed a more potent LAK activity than conventional LAK cells and CD3-activated T cells. These results suggest that selective expansion and activation of CD16 and CD56 positive cells in LAK cells is a useful strategies to improve their anti-tumor potential in nonspecific immunotherapy, and possibly in combination therapy with other target immunotherapies as well.     

OK432治疗脑胶质瘤研究进展

OK43 2 (picibanil)及OK43 2激活的外周血单核细胞 (OK 43 2 activatedmononuclearcells ,OK MCS)作为免疫赋活剂 ,具有较强的抗肿瘤活性。选择手术切除并经过放、化疗的脑胶质瘤患者 ,采取瘤腔内注射给药 ,可激活T细胞、LAK细胞、嗜中性粒细胞及巨噬细胞等 ,并可诱发出细胞毒因子 (tumornecrosisfactor ,TNF) ,从而产生杀伤瘤细胞的作用。基因水平研究亦显示有瘤细胞凋亡的发生。临床应用不良反应轻微、患者耐受性好 ,并可减轻或对抗放、化疗引起的白细胞减少及骨髓抑制等不良反应 ,具有一定的临床效果 ,可作为脑胶质瘤治疗的一种方法。     

Lymphokine-activated killer function following autologous bone marrow transplantation for refractory hematological malignancies

Disease recurrence remains the major factor which limits the success of autologous bone marrow transplantation (ABMT) for refractory hematological malignancies. The administration of interleukin 2 (IL2) with or without ex vivo generated lymphokine-activated killer (LAK) cells represents a potential approach to eradicating residual disease after ABMT. However, since LAK precursor activity is radiosensitive, high dose chemoradiotherapy may abrogate LAK function and preclude clinical responsiveness to IL2 after ABMT. Furthermore, since lymphocyte subsets which mediate LAK activity may recover at different rates after ABMT, LAK cells may be phenotypically and/or functionally altered after ABMT. To determine whether IL2 responsive LAK precursor cells are present in the circulation after ABMT, peripheral blood mononuclear cells (PBMC) from 21 patients with acute leukemia or lymphoma were tested for IL2-inducible LAK activity 17-83 days after ABMT. Cells were cultured with IL2 (1000-2000 units/ml) for 4 or 5 days and then tested for cytolytic activity and/or cell phenotype. LAK activity against the Daudi cell line was detected in every PBMC sample from every patient at every time point tested. The Raji cell line and a fresh allogeneic ovarian carcinoma were also lysed by LAK cells generated after ABMT. In the subgroup of patients transplanted for non-Hodgkin's lymphoma, LAK precursor activity appeared comparable to that of healthy controls. Culture with IL2 resulted in increased mean IL2 receptor expression in lymphocytes from patients after ABMT (3.1-9.9%) and from healthy controls (3.1-12.0%). After culture with IL2, the percentage of cells bearing the natural killer cell-associated Leu-19 determinant was significantly higher in patient PBMC than in normal control PBMC (28.3 versus 8.7%). Positive and negative cell selection by fluorescence sorting after culture with IL2 revealed that most of the LAK activity after ABMT was mediated by the Leu-19+ cells. Although CD5+ T-cells were devoid of LAK activity, a subset LAK effectors was CD8+. Thus, LAK activity is rapidly reconstituted after ABMT and is mediated by cells phenotypically similar to those in normal controls. These results support the feasibility of IL2 +/- LAK as consolidative immunotherapy after ABMT.     

Intrapleural administration of OK432 in cancer patients: Activation of nk cells and reduction of suppressor cells

Twelve patients with carcinomatous pleural effusions were treated with single intrapleural (i.pl.) administration of OK432 on day 0 and the effects of i.pl. OK432 on natural killer (NK) cell activity were followed on day 7. Two patients showed no clinical evidence of therapeutic benefit from i.pl. OK432. In the other 10 patients, pleural effusions and/or tumor cells in the effusions had decreased or disappeared by day 7. NK cell activity was markedly low or absent in pleural effusions of untreated patients due to the presence of adherent effusion cells capable of suppressing the maintenance and interferon-induced augmentation of NK cell activity. I.pl. injection of OK432 resulted in enhancement of NK cell activity and abrogation of NK suppressor cell activity in the effusions. On the other hand, blood NK cell activity was not consistently altered by i.pl. OK432. In vitro treatment of effusion mononu-clear cells from untreated patients with OK432, but not with interferon, augmented NK cell activity. In addition, adherent effusion cells of untreated patients lost their NK suppressor function following in vitro OK432 treatment. These results suggest that i.pl. administration of OK432 will result in augmentation of NK cell activity and reduction of NK suppressor cell activity in pleural effusions, which could be responsible for the antitumor activity of i.pl. OK432.     

CIK细胞体内外抗宫颈癌HeLa细胞活性的研究

目的：探讨细胞因子诱导的杀伤（cytokine-induced killer,CIK）细胞的体内外抗宫颈癌HeLa细胞活性。方法：收集8名健康献血者和8名宫颈癌患者的新鲜外周血,分别通过常规方法分离外周血单核细胞（peripheral blood mononuclear cells,PBMC）,应用相应细胞因子体外诱导分化出CIK细胞,动态观察CIK细胞的体外增殖活性、细胞表型和对HeLa细胞的杀伤活性;在BALB/c裸鼠皮下接种效应细胞,观察宫颈癌患者CIK细胞对接种HeLa细胞的荷瘤鼠的抑瘤作用,同时设淋巴因子激活的杀伤细胞（lymphokine activated killer cells,LAK）和PBMC细胞作为对照。结果：源于健康人和宫颈癌患者的CIK细胞间的增殖活性无明显区别（P〉0.05）。表型分析结果表明,两种来源的CIK细胞中CD3~＋CD56~＋双阳性细胞均得到了大量扩增,宫颈癌患者CIK细胞中CD3~＋CD56~＋双阳性细胞在实验开始前约占0.13%,到实验后第28天上升到25.8%。体外实验表明,宫颈癌患者的CIK细胞杀伤宫颈癌HeLa细胞的细胞毒活性明显高于PBMC细胞。裸鼠体内实验表明,宫颈癌患者CIK细胞能够显著抑制肿瘤的生长,其抑瘤率可达80.6%,高于LAK细胞的59.1%和PBMC细胞的38.3%（P〈0.01）。CIK治疗后肿瘤体积明显比空白对照组缩小（P〈0.05）。结论：宫颈癌患者CIK细胞具有较强的体内外抗宫颈癌细胞活性,有可能用于临床上宫颈癌的过继性免疫治疗。     

Cytotoxicity of purified Leu 19+ cells from the peripheral blood of children with acute lymphocytic leukemia.

Several investigators have reported decreased natural killer cell activity of the peripheral blood from children with acute leukemia both at diagnosis and in relapse. This study was to determine whether this decreased natural killer activity is the result of a diminished number or defective function of natural killer cells or both. The percentage of Leu 19+ (NKH-1, CD56) and the lytic activity of a purified Leu 19+ population from the peripheral blood of children with acute lymphocytic leukemia as well as from healthy children were determined. This cell population mediates most natural killer and natural killer-like activity in the peripheral blood. Twenty-three children with acute lymphocytic leukemia (16 at diagnosis, seven in relapse) as well as six control children were studied. The percentage of Leu 19+ cells was significantly lower in the blood of children either at initial diagnosis (median, 3%; P less than 0.005) or in relapse (median, 1%; P less than 0.01) than that for the control children (median, 12.5%). The cytotoxicity of peripheral blood mononuclear cells (PBMC) against K562 in 3-hour 51Cr release assay was significantly lower for patients either at diagnosis (median, 8.5 lytic units[LU]/10(7) cells; P less than 0.005) or in relapse (median, 3 LU/10(7) cells; P less than 0.005) than that for normal controls (median, 97 LU/10(7) cells). The lytic activity of a fluorescent activated cell sorter (FACS) purified Leu 19+ population was evaluated. The Leu 19+ cells were sorted from PBMC of 11 children with acute lymphocytic leukemia who had more than 2% Leu 19+ cells in their PBMC (nine at diagnosis and two in relapse). The lytic activity of sorted Leu 19+ cells was significantly (P less than 0.04) lower for patients at initial diagnosis (median, 45 LU/10(7) cells) than that for normal control children (median, 317 LU/10(7) cells). One of the two children studied at relapse had low Leu 19+ lytic activity. The authors concluded that (1) The percentage of Leu 19+ cells in the peripheral blood of the majority of children with acute lymphocytic leukemia either at diagnosis or in relapse is below normal; (2) The lytic activity of PBMC from the majority of children with acute lymphocytic leukemia is reduced both at diagnosis and in relapse; and (3) The lytic activity of Leu 19+ purified PBMC is reduced in the majority of children with acute lymphocytic leukemia. These findings confirm the defective natural killer cell profile in children with acute lymphocytic leukemia both at the time of diagnosis and in relapse.     

The heterogeneity of target recognition by lymphokine-activated killer precursor cells

Lymphokine-activated killer (LAK) cells were generated from peripheral blood lymphocytes (PBL) that were depleted of mature cytotoxic natural killer (NK) cells. PBL NK activity was abolished by pretreatment of effector cells with the toxic lysosomotropic agent L-leucine methyl ester (LME) or by depletion of effector cells by K562 monolayer absorption (MA). Both treatments markedly reduced the proportion of cells expressing NK-associated markers such as CD 16 (Leu 11b, B73.1), Leu 7, and NKH-1 (Leu 19), whereas these treatments had minimal effects on cells expressing T cell markers (CD 3, CD 4, and CD 8). LME and MA also drastically decreased the proportion of K562 target-binding lymphocytes. LAK activity against NK-sensitive and NK-resistant targets can be generated from the NK cell-depleted PBL by incubation with interleukin-2. Peak LAK activity generated from MA-treated PBL was later than the peak of LAK activity generated from either untreated or LME-treated PBL. Although MA of PBL on NK-resistant S4 sarcoma targets had little effect on NK activity, LAK activity against both K562 and S4 targets was reduced. These results suggest that there are at least three LAK precursor subpopulations in PBL: mature NK cells that can bind and kill K562 targets (LME-sensitive and MA-sensitive); "pre-NK" cells that can bind but cannot kill (LME-resistant and MA-sensitive); and non-NK cells that cannot bind and cannot kill K562 targets (MA-resistant).     

脂质体介导的基因转染效率与肺癌细胞的增殖特性的关系

目的：简化粘附性LAK（A－LAK）细胞的制备方法并观察其与苯乙酸（PA）的协同抗瘤作用。方法：采用苯丙氨酸甲酯（PME）处理肝癌患者外周血单个核细胞（PBMC）而制备A－LAK细胞;观察人肝癌SMMC7721细胞株经PA处理后增殖能力的变化;并用经PA预处理的SMMC7721培养上清液作用于A－LAK细胞,观察A－LAK增殖能力与杀伤活性的变化。结果：采用PME制备的A－LAK细胞,其增殖能力显著高于非粘附性LAK细胞（NA－LAK）和常规LAK细胞;肿瘤细胞经PA作用后,生长明显受 到抑制;肿瘤细胞的培养上清液可明显抑制A－LAK的增殖与杀伤活性,而经苯乙酸预处理的上清液则抑制作用减弱。结论：采用PME可 简便快速地制备A－LAK,而苯乙酸可与A－LAK协同发挥抗瘤作用。     

Inhibition of lymphokine-activated killer cells generation in vitro by soluble factors released from mixed human tumor and peripheral blood mononuclear adherent cells culture

Background: A variety of molecules produced by both tumor cells and normal cells reduce the activity of lymphokine-activated killer (LAK) cells. We tested the possible cross-regulation of mel-624 melanoma cells and adherent peripheral blood mononuclear cells (PBMCs) in affecting LAK cell activity. Methods: PBMC adherent cells were cultured together with mel-624 melanoma cells. Supernatant was transferred to a 4-day LAK cells generation culture consisted of PBMC nonadherent cells and interleukin-2. LAK cytotoxic activity was tested in a 4-hour assay against Daudi tumor cells prelabeled with sodium ⁵¹chromate. Results: The supernatant produced within the first 48 hours of mixed mel-624 melanoma cell and adherent PBMC culture substantially (by 69 percent) reduced the generation of LAK cells, whereas the supernatant from either tumor culture or adherent PBMC culture had no effect. The inhibitory effect was manifested on the generation of LAK cells when autologous nonadherent cells were cultured with 1,000 units/ml IL-2, but there was no effect on mature LAK cell cytotoxic activity. Inhibition of LAK cell generation was partially dependent on protein synthesis and was not mediated by transforming growth factor β (TGF-β). Conclusion: Our results point toward the production of soluble, yet unidentified proteins, in mixed tumor-adherent PBMC cultures, which substantially reduced the induction of LAK cells in culture.     

The Heterogeneity of Target Recognition by Lymphokine-activated Killer Precursor Cells

Lymphokine-activated killer (LAK) cells were generated from peripheral blood lymphocytes (PBL) that were depleted of mature cytotoxic natural killer (NK) cells. PBL NK activity was abolished by pretreatment of effector cells with the toxic lysosomotropic agent l -leucine methyl ester (LME) or by depletion of effector cells by K562 monolayer absorption (MA). Both treatments markedly reduced the proportion of cells expressing NK-associated markers such as CD 16 (Leu 11b, B73.1), Leu 7, and NKH-1 (Leu 19), whereas these treatments had minimal effects on cells expressing T cell markers (CD 3, CD 4, and CD 8). LME and MA also drastically decreased the proportion of K562 target-binding lymphocytes. LAK activity against NK-sensitive and NK-resistant targets can be generated from the NK cell-depleted PBL by incubation with interleukin-2. Peak LAK activity generated from MA-treated PBL was later than the peak of LAK activity generated from either untreated or LME-treated PBL. Although MA of PBL on NK-resistant S4 sarcoma targets had little effect on NK activity, LAK activity against both K562 and S4 targets was reduced. These results suggest that there are at least three LAK precursor subpopulations in PBL: mature NK cells that can bind and kill K562 targets (LME-sensitive and MA-sensitive); "pre-NK"cells that can bind but cannot kill (LME-resistant and MA-sensitive); and non-NK cells that cannot bind and cannot kill K562 targets (MA-resistant).     

Induction of activated natural killer cells by the streptococcal preparation OK432 (I). Induction from spleen cells of normal or tumor-bearing mouse primed in vivo and subsequently challenged in vitro with OK432

The present study shows that natural killer cell-mediated cytotoxicity of BALB/c mouse spleen cells to syngeneic tumor cells was augmented by in vivo priming or in vitro stimulation with the streptococcal preparation OK432. The augmentation of spleen cell cytotoxicity to syngeneic tumor cells by in vivo priming alone with OK432 was lower than that obtained by in vitro stimulation alone with OK432. When the murine spleen cells primed in vivo with OK432 were rechallenged in vitro with OK432 at various intervals, the natural cytotoxicity was more strongly enhanced than that seen with in vitro stimulation alone. The cell surface phenotype of killer cells activated with OK432 was Thy1+ and asialo GM1+, suggesting the activated natural killer cell. Mice were transplanted with syngeneic colon adenocarcinoma cells, and primed in vivo with OK432, and then their spleen cells were subsequently challenged in vitro with OK432. These cells displayed a strong cytotoxic activity not only to the transplanted adenocarcinoma cells but also to other syngeneic tumor cells.     

Identification of a novel CD56- lymphokine-activated killer cell precursor in cancer patients receiving recombinant interleukin 2.

Circulating lymphokine-activated killer (LAK) cell activity in cancer patients receiving recombinant interleukin 2 (rIL-2) therapy is confined to cells expressing the CD56- surface marker. However, CD56- cells from these patients but not normal individuals have been reported to exhibit LAK cytotoxicity only following in vitro activation with rIL-2. Studies were performed to document the existence of CD56- LAK precursor cells and to phenotypically characterize this population in patients receiving rIL-2 therapy using fluorescence-activated cell sorter-purified CD56- cell subsets. Initial studies confirmed that CD56- cells exhibit NK activity [20 +/- 7 (SE) LU/10(6) cells] but not LAK activity (0 +/- 0 LU/10(6) cells) when evaluated directly from peripheral blood of patients receiving rIL-2. CD56- cells from patients but not normal individuals developed significant LAK cytolytic activity against NK-resistant COLO 205 targets (16 +/- 3 LU/10(6) cells) when cultured for 3 days with 1500 units/ml rIL-2. The CD56- LAK precursor activity was confined to cells expressing a CD56-CD16+ phenotype and a large granular lymphocyte morphology; little or no NK or LAK precursor activity was detectable in CD56-CD5+ T-cells from patients. Phenotypic characterization of CD16+CD56- cells revealed that this population is uniformly CD11a+,CD18+, and CD38+ and is heterogeneous in its expression of CD11b, CD11c, and CD16/Leu 11c. These results indicate that rIL-2 administration induces enhanced LAK precursor activity in a novel population of CD5-CD16+CD56- cells.     

Natural killer and lymphokine-activated killer activities in stomach cancer patients with special emphasis on the effect of 5-fluorouracil, adriamycin and mitomycin-C chemotherapy

The cytotoxicities of peripheral blood lymphocytes (PBL) and lymphokine-activated killer (LAK) cells were studied to evaluate the effect of chemotherapy on cellular immunity, in 18 patients with unresectable stomach cancer before and after chemotherapy with 5-fluorouracil, adriamycin and mitomycin-C (FAM), and in 21 healthy volunteers. LAK cells were generated in vitro by culturing PBL with 100 U recombinant human interleukin-2 (rH-IL-2)/ml for 72 h. K562 (human myelogenous leukemia), MKN-45 (human stomach adenocarcinoma) and PC-14 (human pulmonary adenocarcinoma) were used as target cells. The cytotoxicity of PBL to K562 and MKN-45 was suppressed in patients with stomach cancer before chemotherapy, compared with that in healthy volunteers (P less than 0.05). The cytotoxicity of LAK cells was significantly higher to all three cell lines tested than that of PBL in both the healthy volunteers and stomach cancer patients (P less than 0.01); however, a lower level of LAK activity was generated in patients with cancer compared to that in the healthy volunteers. FAM therapy did not suppress the cytotoxicities of PBL and LAK cells. The surface markers of PBL and LAK cells were measured, demonstrating that there was no significant change in the percentage of lymphocytes with CD3+, CD4+, CD8+, CD16+ or CD19+ after chemotherapy. The ratios of CD4+ to CD8+ cells in PBL and LAK cells were also not significantly changed after chemotherapy. In the present study, we have demonstrated that the PBL of stomach cancer were defective in generating LAK activity compared to those of controls, but the LAK activity generated from PBL receiving chemotherapy was similar to that from PBL without chemotherapy in stomach cancer patients.     

Generation of human lymphokine-activated killer cells following brief exposure to high dose interleukin 2

Current laboratory lymphokine-activated killer (LAK) cell activation procedures require culture of peripheral blood mononuclear cells (PBMC) in the presence of 1000-1500 units/ml of interleukin 2 (IL-2) for 3-7 days. However, we have observed that a brief exposure (15 min-1 h) of PBMC to a high concentration of IL-2 results in the maturation of LAK precursor cells to cytolytic effector cells over the course of 1-3 days. These IL-2-pulsed LAK cells express cytolytic activity comparable to that of nonpulsed PBMC (cultured continuously in IL-2) at 3 days of culture. The acquisition of cytolytic activity followed the same kinetics for both pulsed and nonpulsed mononuclear cells and was maintained when tested at day 7. The pulsed LAK cells were capable of significantly lysing 11 different tumor targets tested and flow cytometric analysis revealed that pulsed LAK cells were phenotypically similar to nonpulsed LAK cells. Serum obtained from cancer patients undergoing IL-2/LAK cell therapy did not inhibit the maturation of the pulsed mononuclear cells into LAK cells. Interestingly, only PBMC obtained from cancer patients receiving in vivo IL-2 infusions could be induced to generate the same levels of cytolytic activity as those in nonpulsed cells using this pulse procedure. PBMC obtained from healthy, normal donors could not be pulsed to the same levels of activation as nonpulsed LAK cultures. Our study demonstrates that for the generation of maximum LAK cell cytolytic activity, LAK cell precursors must be primed in vivo with IL-2. Implementation of this procedure could eliminate the high cost of cell culture which normally accompanies IL-2/LAK cell therapy. Such an approach could make IL-2/LAK cell therapy more accessible for cancer patients.     

脐血CD_3AK细胞的制备、生物活性测定及临床应用初探

 目的　研究用脐血单个核细胞制备CD3 AK细胞的抗肿瘤作用 ,探讨肿瘤生物治疗近期疗效的免疫指标。方法　分离脐血单个核细胞 ,分别用IL 2和IL 2 +CD3 Ab诱导LAK和CD3AK细胞 ,并测其扩增数量和对K5 6 2细胞的杀伤活性 ;测定肿瘤患者用CD3 AK细胞治疗前后外周血单核细胞(PBMC)的NK杀伤活性。结果　脐血LAK细胞和脐血CD3 AK细胞均于培养后第 11天时扩增倍数最高 ,分别是培养前的 18倍、2 4倍 ,对K5 6 2的杀伤活性分别是培养前的 2 .6倍和 3.2倍 ;肿瘤患者输注CD3 AK细胞一疗程后 ,其PBMC的NK杀伤活性由 6 2 %升高到 82 % ,平均升高 32 %。结论　 (1)脐血单个核细胞是LAK细胞和CD3 AK细胞良好的前体细胞。 (2 )LAK和CD3 AK细胞的数量和杀伤能力在培养的第 11天时达高峰 ,而且CD3 AK细胞数量和杀伤活性明显优于LAK细胞。 (3)CD3 AK细胞输注能明显提高肿瘤患者PBMC的NK活性。 (4 )肿瘤病人PBMC的NK活性测定可望成为肿瘤生物治疗的一个近期疗效参数     

Immunotherapy of terminal-stage malignant tumors with the immunopotentiator OK-432--a comparison between SU-PS test-responder and -nonresponder patients

We treated terminal malignant tumor cases with the immunopotentiator OK-432 and determined the absolute numbers of neutrocytes, lymphocytes, and lymphocyte subpopulations, carrying out the SU-PS and PPD skin tests. After the start of therapy, SU-PS test changed to positive in one-third of the patients. The SU-PS -responder patients showed an increase in lymphocytes and Leu 11A-positive cells, and an elevation of the OK T4/T8 ratio two weeks later. In the SU-PS-nonresponder group, OKT4-positive cells declined, while OKT8-positive cells increased with time. That is, the OKT4/T8 ratio remained low throughout the test period. In the SU-PS-responder group, OK-432 therapy was found to prolong the survival period.