Locoregional adoptive immunotherapy using LAK cells and IL-2 against liver metastases from digestive tract cancer

We investigated the clinical efficacy of locoregional and systemic adoptive immunotherapy (AIT) with or without interleukin-2 (IL-2) against solid metastatic lesions from digestive tract cancer. Eighteen patients, who were treated with more than 10(10) lymphokine-activated killer (LAK) cells, were enrolled in this study. Seven of the 18 patients received hepatic arterial infusion (HAI) of LAK cells with or without IL-2 against metastatic liver tumors (locoregional therapy group). The remaining 11 patients received systemic transfer of LAK cells with IL-2 against metastatic lesions located in organs other than the liver (systemic therapy group). Three of 7 locoregional therapy group patients showed clinically significant tumor regressions that were evaluated as being equivalent to partial response (PR). Two of the 11 systemic therapy group patients showed significant tumor regressions, but this response rate was much lower than that of the locoregional therapy group. The 2 effective cases in the systemic therapy group were esophageal cancer patients. Locoregional AIT with or without IL-2 against liver metastases from digestive tract cancer could be an effective therapeutic modality in some patients who are refractory to conventional therapies (e.g., chemotherapy and radiotherapy). It is necessary to find a new way to augment the anti-tumor effect of this therapy in combination with prior or concomitant chemotherapy.     

Locoregional adoptive immunotherapy resulted in regression in distant metastases of a recurrent esophageal cancer

 Esophageal cancer is one of the most common malignant diseases. However, postoperative recurrences are still resistant to currently available radiochemotherapy. We recently reported a study on the initial clinical efficacy of locoregional adoptive immunotherapy for advanced esophageal cancer. We report here our clinical experience of remarked responses in distant metastatic lesions in a patient with recurrent cancer after receiving this immunotherapy. A male patient underwent curative surgery, and presented with multiple recurrent metastases in the supraclavicular lymph nodes (LNs), liver, and abdominal aortic LNs. Autologous tumor-activated lymphocytes (AuTLs) generated ex vivo were regionally injected into supraclavicular LNs every 2 weeks 13 times. Mean numbers of the administrated cells were 0.8 × 10⁹ cells/injection. AuTLs established from peripheral blood lymphocytes stimulated by autologous tumor cells with interleukin-2 were tested for their cytotoxicity before every treatment. During immunotherapy, Grade 2 diarrhea and fever were observed. The clinical partial responses were obtained in all lesions and were sustained for 11 months. Because clinical toxicity was tolerable, this immunotherapy might be useful for patients with far-advanced esophageal cancers. Received: June 19, 2002 / Accepted: September 26, 2002 Correspondence to:U. Toh     

Intraarterial cellular immunotherapy for patients with inoperable liver metastases of esophageal cancer

PURPOSE: We report 2 patients with refractory liver metastatic tumor after esophagectomy for advanced esophageal cancer, who responded markedly to locoregional cellular immunotherapy by repeated intraarterial infusions of autologous tumor cell-activated T lymphocytes (AuTL), even after they failed the standard chemotherapy of cisplatin (CDDP) and 5-fluorouracil (5-FU). METHODS: AuTL administrations were made through the hepatic artery via a subcutaneous reservoir located at the right upper leg. Six injections were administered to both patients, repeated at 2-week intervals. The total number of administered T cells reached 2.4 x 10(9) and 3.1 x 10(9), respectively. RESULTS: A 39% and 51% regression in each infused field, compared with the size of liver tumor before treatment, was observed by computed tomography (CT) scan in patient 1 and 2, respectively. The responses continued up to the 10th and 11th month after the intraarterial infusion, confirmed by follow-up CT scan. The adverse effects of intraarterial immunotherapy were tolerable, with grade 1-2 fever and nausea in each patient. CONCLUSIONS: Clinical regression of liver metastases of esophageal cancer was observed in both patients who received this intraarterial cellular immunotherapy. Liver metastases of esophageal cancer may be controlled effectively and safely by repeating the intraarterial AuTL infusion as a locoregional immunotherapy over a long period.     

Combination immunotherapy using autologous tumor-stimulated lymphocytes and trastuzumab (Herceptin) for the patients with recurrent breast cancer

PURPOSE: We report two patients with refractory recurrent breast cancer (HER2/neu: +) postoperatively, who had failed response to the available conventional chemotherapy of CAF (cyclophosphamide, adriamycin, 5-fluorouracil) and docetaxel, etc. They markedly responded to the combination immunotherapy using intraperitumoral injections of autologous tumor cell-stimulated T lymphocyte (AuTL) and trastuzumab (Herceptin), an anti-HER2 monoclonal antibody. METHODS: AuTLs were administrated directly into the recurrent tumor by intraperitumoral injections biweekly and trastuzumab was infused systemically every week. The treatments were repeated for 6 and 11 injections in the patients, respectively. The total administered T cells had reached to 3.8 x 10(9) and 6.4 x 10(9), respectively. The dosage of trastuzumab was 2 mg/kg in each patient. RESULTS: The carcinomatous pleural effusion had disappeared and was well controlled in patient 1 and a marked regression in injected fields in comparison to the size of the recurrent tumor before treatment was observed in patient 2. The tumor marker proteins (CEA, CA15-3, TPA) had also decreased significantly. The adverse effects of the immunotherapy were tolerable with grades 1-2 infusion reaction of fever, tachycardia and hypotension, but no cardiac dysfunction was observed. CONCLUSIONS: Clinical responses of recurrent breast cancer were observed in two patients after receiving the intra-peritumoral AuTL injection plus trastuzumab immunotherapy. These results showed that refractory recurrent breast cancer may be controlled effectively and safely by repeating the cellular immunotherapy combined with trastuzumab and suggested utility of combining these agents in clinical trial.     

33例头颈部恶性肿瘤患者局部过继免疫治疗的疗效观察

目的评价ＩＬ－２／ＬＡＫ细胞局部过继免疫疗法在头颈部恶性肿瘤治疗中的疗效。方法对３３例头颈部恶性肿瘤患者进行局部过继免疫治疗,采用ＩＬ－２每日１０～２０万单位局部注射,共１０天;于ＩＬ－２治疗的第４～８天同时于局部注射ＬＡＫ细胞１．０×１０８～５．０×１０８／ｄ。结果完全缓解１例,部分缓解６例,好转２０例,稳定６例,治疗总缓解率２１．２％,总有效率８１．８％。治疗后１,２,３年生存率分别为９６．３％、８３．３％、和７５．０％。组织病理学检查证实免疫治疗后肿瘤局部大量ＣＤ３、ＣＤ４阳性Ｔ淋巴细胞浸润。治疗过程中未见严重的毒副作用。结论局部应用ＬＡＫ细胞与ＩＬ－２治疗头颈部恶性肿瘤疗效明显,方法安全。     

Local adoptive immunotherapy using lymphokine-activated killer cells and interleukin-2 against malignant pleural mesothelioma: report of two cases.

Malignant pleural mesothelioma is refractory to conventional therapy. We tried local adoptive immunotherapy using lymphokine-activated killer (LAK) cells and interleukin-2 (IL-2) to control malignant pleural effusion due to mesothelioma in two patients: Case 1 was that of a 69-year-old man, and Case 2 that of a 49-year-old woman with complicating chronic idiopathic thrombocytopenic purpura. A systemic survey revealed no sign of metastasis in either case. Intrapleural instillations of 6.7 x 10(9) autologous LAK cells in Case 1, or 11.3 x 10(9) allogeneic LAK cells in Case 2, with daily injections of IL-2 resulted in reductions of the pleural effusions in each case and in a decline in the level of hyaluronic acid in the effusion, in Case 1. The instillations of autologous and allogeneic LAK cells were well tolerated. The results suggest that local adoptive immunotherapy could be useful in the treatment of malignant effusion due to mesothelioma.     

Different immune functions of peripheral blood, regional lymph node, and tumor infiltrating lymphocytes in lung cancer patients

Immune functions of peripheral blood (PBL), regional lymph node (RLNL), and tumor infiltrating lymphocytes (TIL) were evaluated in lung cancer patients. PBL had many natural killer (NK) cells and the highest NK activity, and it showed the highest augmentation of NK activity by interferon-gamma (IFN-gamma) + recombinant interleukin-2 (rIL-2) among the three groups of lymphocytes. PBL had high lymphokine-activated killer (LAK) activity of against a broad spectrum of cell lines and moderate activity against autologous tumor cells by increased effector to target (ET) ratio but the lowest ability of IL-2 production of the three groups of lymphocytes. The RLNL not associated with tumor metastasis had a few NK cells and lower NK activity than PBL, but its LAK activity was almost the same but not greater than that of PBL. RLNL had the highest ability of IL-2 production among the three groups of lymphocytes. All activities of RLNL associated with tumor metastasis were lower than those not associated with tumor metastasis. TIL exclusively consisted of T-cells, especially cytotoxic/suppressor T-lymphocytes. NK activity and lymphocyte blastogenesis of TIL were lower than those of other groups. The LAK activity of TIL differed greatly with the case, and it was the highest against autologous tumor cells among the three groups of lymphocytes in three of eight cases. These findings showed that PBL, RLNL, and TIL had characteristic subpopulations of lymphocytes and different functions of host immune responses in lung cancer. Efficient augmentation of the characteristic immune responses will lead to a more effective total cancer therapy.     

肺癌肿瘤全溶物脉冲自体树突状细胞体外诱生T细胞抗肿瘤反应的研究

目的　探讨非小细胞肺癌 (NSCLC)患者全瘤溶解物 (WTL)冲击的自体树突状细胞 (DCs)瘤苗体外诱生T细胞介导的抗肿瘤反应。方法　在含重组人粒细胞 巨噬细胞 集落刺激因子 (rhGM CSF)和重组人白细胞介素 4 (rhIL 4 )的培养条件下 ,以 10例患者的贴壁外周血单核细胞 (PBMCs)衍生不成熟的树突状细胞 (imDCs) ,分别添加肿瘤坏死因子 (TNFα)、自体WTL或WTL TNFα诱导成熟 ,即分别为DCs/TNF、DCs/WTL和 DCs/WTL ,用流式细胞仪和混合淋巴细胞反应 (MLR) ,检测细胞表面分子的表达变化及其刺激异基因或同基因的T细胞增殖能力。将 DCs/WTL和自体T细胞共培养1～ 2周 ,以诱导细胞毒性T淋巴细胞 (CTL) ,用计数γ 干扰素 (IFN γ)释放的酶联免疫斑点 (ELISPOT)试验和乳酸脱氢酶 (LDH)释放的细胞毒试验 ,分别检测该CTL中识别自体肿瘤细胞释放特异性IFN γ的T细胞数和溶瘤活性。结果　以 30 μg/ml蛋白的WTL体外冲击自体 10 6imDCs,可导致上调DC表型 ,包括CD1a、CD83和CD86以及HLA DR的分子表达 ,与常规TNFα诱导成熟的DCs表型相似。虽然两者均能显著刺激异基因T细胞的增殖 ,但其刺激自体T淋巴细胞的增殖能力 ,DCs/WTL却显著高于DCs/TNF(P <0 .0 5 )。将体外与 DCs/WTL共培养的自体T细胞作为反应或效应细胞 ,再次     

Adoptive immunotherapy of human cancer using low-dose recombinant interleukin 2 and lymphokine-activated killer cells

The adoptive transfer of recombinant-methionyl human interleukin 2 (rIL-2)-activated autologous peripheral blood mononuclear lymphokine-activated killer (LAK) cells to cancer patients is being evaluated as an alternative to conventional cancer therapy. We have independently developed an alternative regimen to previously reported adoptive immunotherapy protocols using rIL-2 and LAK cells which features the prolonged administration of low-dose rIL-2 (30,000 units/kg) and an automated, entirely enclosed system of peripheral blood cell procurement, culture, harvest, and reinfusion of activated cells. The cell culture system was tested with a murine tumor model in which LAK cells generated in plastic culture bags were reinfused into tumor-bearing mice. Tumor regression was as effective with cells activated in the bags as in conventional culture flasks. Twenty-eight cancer patients were treated for 5 consecutive days with low-dose rIL-2, followed by leukapheresis, infusion of LAK cells, and prolonged IL-2 administration. At least 50% tumor regression was observed in 46% of all patients treated. These data imply that human peripheral blood mononuclear cells retain fully their capacity for rIL-2-induced activation and effector cell function under this alternative approach, and further, that a low-dose rIL-2 regimen with markedly reduced toxicities can be as effective as high-dose rIL-2 regimens if low-dose rIL-2 is given for a prolonged period of time following LAK cell infusion.     

Natural killer and lymphokine-activated killer cell-mediated cytotoxicity in patients with oral cancer

Peripheral blood lymphocytes (PBL) from untreated and treated oral cancer patients, lymph node lymphocytes (LNL) from metastatic (met) and nonmetastatic (non-met) lymph nodes, and tumor infiltrating lymphocytes (TIL) were tested for natural killer (NK) and lymphokine activated killer (LAK) cell cytotoxicity using appropriate targets in a short-term chromium release assay. The results showed that while both NK and LAK functions of PBL from oral cancer patients were comparable to those of normal healthy donors, the NK activity of metastatic and nonmetastatic LNL and TIL was highly compromised. On the other hand, potent LAK activity could be generated from all three lymphoid populations. Individual patients showing low NK activity displayed good LAK cytotoxicity, indicating that endogenous cells with low NK potential have adequate ability to respond to interleukin 2 (IL-2). LAK activity tested on autologous tumour targets revealed that TIL were the best source of LAK cells. followed by PBL and LNL.     

Induction of tumor-cell lysis by bi-specific antibody recognizing ganglioside GD2 and T-cell antigen CD3

Human tumor cells expressing ganglioside GD2 were lysed by various effector populations targeted with an anti-CD3-anti-GD2 bi-specific antibody (BAb CD3 x GD2). This antibodyheteroconjugate was prepared by chemically cross-linking the OKT-3 monoclonal antibody (MAb) reactive with CD3 antigen on T lymphocytes with the ganglioside MAb ME 361, which binds preferentially to the tumor-associated ganglioside GD2. The specificity of target-cell lysis by the cytotoxic T cells (CTL) was mediated by the specificity of the targeting antibody: GD2-negative cells were not lysed in the presence of the CD3 x GD2 BAb. A dose-dependent response was observed in a range of 10 to 10,000 ng/ml. In contrast, 2 other BAbs recognizing the tumor-associated antigens EGF-R and TKB-2 had greater potency to mediate tumor-cell lysis than the GD2 x CD3 BAb. Peripheral-blood cells (PBL) stimulated with OKT-3 MAb or with irradiated tumor cells in a mixed lymphocyte culture (MLTC) could be induced to lyse GD2-positive tumor cells in the presence of CD3 x GD2 BAb. The tumor-cell lysis could be mediated by autologous or allogeneic effector cells. NK cells had no influence on the BAb-induced cytotoxicity.     

Lymphokine-activated killer-cell function of lymphocytes from peripheral blood, regional lymph nodes and tumor tissues of patients with oral cancer

Lymphokine-activated killer (LAK) cells, generated from peripheral blood lymphocytes (PBL) from patients with oral cancer or oral leukoplakia and from healthy donors showed comparable lysis of 6 target tumor cell lines, including 3 derived from head and neck and oral cancers. The tumor burden of the host did not appear to influence the systemic LAK activity. LAK activity of lymphocytes infiltrating the tumor tissues (TIL) was also comparable to that of the PBL. Both TIL and PBL showed a parallel increase in proportion of HNK-I+ and CD-25+ cells upon activation with IL-2. The lymph-node lymphocytes (LNL) from metastatic (met) and non-metastatic (non-met) draining lymph nodes, however, showed reduced LAK activity and an increase in CD8+ cells, in addition to CD25+ and HNK-I+ cells, when cultured with IL-2. When IL-2-activated LNL were co-cultured with autologous PBL during IL-2 activation of the latter, a strong suppressive effect was exerted by LNL. In contrast, IL-2-activated PBL did not suppress autologous LAK generation in spite of an increase in CD8+ cells seen after activation with IL-2. Frequency distribution of LAK precursors was significantly lower in LNL than in PBL from oral cancer patients. LAK precursor frequency in TIL was comparable to that of PBL. The results show that, in oral cancer, regional lymph nodes may not have adequate IL-2-inducible cytotoxic potential, due to a reduced number of LAK progenitors and possible activation of suppressor cells. Alternatively, TIL can be a potential source for LAK cell function.     

Lymphokine-activated killer (LAK) cells. Analysis of factors relevant to the immunotherapy of human cancer

Lymphokine-activated killer (LAK) cells can be generated by incubating fresh peripheral blood lymphocytes (PBL) in Interleukin-2 (IL-2). LAK cells kill fresh autologous and allogeneic human tumor cells in vitro. This study analyzes aspects of LAK cells that make them a promising candidate for the adoptive immunotherapy of human cancer. LAK cells can be generated from PBL of normal individuals and tumor-bearing patients. Pure, recombinant IL-2 generates LAK cells capable of killing a wide variety of tumors including sarcomas and cancers of the colon, pancreas, adrenal gland, and esophagus. Thirty-six of 41 (88%) fresh, noncultured, human tumor cell suspensions prepared from surgical specimens were lysed by LAK cells in a standard 4-hour chromium-release assay. Normal PBL were not killed. LAK cells can be expanded in vitro for periods longer than 2 months, potentially more than 10(20)-fold, while maintaining lytic ability. These results and the demonstrated efficacy of LAK cells in the therapy of murine tumors make LAK cells a candidate for clinical use in the adoptive immunotherapy of human cancer.     

人肿瘤浸润淋巴细胞（TIL）的抗肿瘤活性及与外周血淋巴细胞...

In order to search for more efficient effector cells than the classical LAK cells for tumor immunotherapy, antitumor activity of TIL was studied. Although fresh TIL were unable to kill both NK-sensitive K562 and NK-resistant Anip 973 targets, rIL-2 activated TIL from 8 lung carcinoma and 4 gastric cancer patients did express significant cytotoxicity against allogeneic solid tumor targets Antitumor activity of activated TIL was more efficient than that of activated autologous PBL By limiting dilution assay, TIL were shown to have a higher frequency of LAK precursors than PBL.     

The repetitive immune cell transfer therapy combining non-myelosuppressive chemotherapy for patients with advanced and refractory cancer

Autologous tumor cells stimulated with T lymphocytes (AuTL) were generated ex vivo from peripheral blood lymphocytes over a two-week co-culturing process with autologous tumor cells. These AuTLs were capable of lysing established tumor cell lines and may have a potential for efficacy as an adoptive immunotherapy (IT) in advanced and metastatic refractory cancer patients (pts). We investigated the feasibility of a combination of AuTL transfer and chemotherapy (ChT) based on the conventional conditioning regimen in order to take advantage by both the anticancer effects and reconstruction of antitumor immunity. Nineteen patients were enrolled in a pilot clinical trial. The two administrations of AuTL were given prior to chemotherapy (ChT) for one treatment cycle. The treatment was repeated at least for three cycles over a one-week interval. The conventional ChT regimen was based on the standard dosage. The pts consisted of 3 of gastric cancer, colon cancer, lung adenocarcinoma, respectively, 6 of esophageal cancer, and 2 of breast and pancreas carcinoma, respectively. AuTLs were administered 1x/2 weeks using direct injection or intraarterial infusion. The median duration of the treatment was over 11.5 months, and the median survival time was 14.8 months. Adverse events related to both the ChT and AuTL transfers at all dosages were minimal. Four of the 13 pts achieved major tumor responses (2 CR: complete regression and 2 PR: partial regression) in this study. Three pts showed progressive disease, and 6 pts had stable disease for over 90 days. PBMC were evaluated for cytokine production prior to the treatment and after 3 treatments. Two and one of 4 CR/PR pts had increased IFN-gamma and TNF-alpha production with no TGF-beta1 responses by their PBMC after 3 treatments, respectively. Two out of 6 pts who experienced stable disease after the treatment had high IFN-gamma and TNF-alpha responses and no TGF-beta1 or IL-4 response. TGF-beta1 and IL-4 secretion increased in parallel in 3 out of 3 pts that experienced progressive disease after the treatment. These data show that combination therapy of AuTL transfer and non-myeloablative ChT is a feasible option for patients with refractory advanced cancers without serious adverse events and without reducing Th1 cytokine responses in peripheral blood for most of the pts that responded to the treatment. According to each mechanism of IT and ChT, a more stringent evaluation of AuTL transfer combined with non-myeloablative ChT for various kinds of cancers should be performed to manage the immunodeficiency in the pts with advanced cancer and to improve the effect of antitumor AuTLs.     

脐血CD3AK细胞治疗恶性肿瘤的临床应用研究

目的:研究用脐血单个核细胞制备CD3AK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 CD3Ab诱导LAK和CD3AK细胞,并测其扩增数量和对K562细胞的杀伤活性;测定肿瘤患者用CD3AK细胞治疗前后外周血T淋巴细胞亚群绝对计数的变化情况和外周血单核细胞(PBMC)的NK杀伤活性.结果:脐血LAK细胞和脐血CD3AK细胞均于培养后11天时扩增倍数最高,分别是培养前的18倍、24倍,对K562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CD3AK细胞一疗程后,其外周血T淋巴细胞亚群绝对计数有明显升高:总T升高66.0%,Th升高68.0%,Ts升高58.0%;其PBMC的NK杀伤活性由63.0%升高到81.0%,平均升高28.0%.结论:1)脐血单个核细胞是LAK细胞和CD3AK细胞良好的前体细胞.2)LAK和CD3AK细胞的数量和杀伤能力在培养的11天时达高峰,而且CD3AK细胞数量和杀伤活性明显优于LAK细胞.3)CD3AK细胞输注能明显提高肿瘤患者外周血T淋巴细胞亚群绝对计数和PBMC的NK活性.4)肿瘤患者的外周血T淋巴细胞计数和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.     

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+.     

Increased susceptibility to lymphokine activated killer (LAK) lysis of relapsing vs. newly diagnosed acute leukemic cells without changes in drug resistance or in the expression of adhesion molecules.

The NK and LAK activity of peripheral blood lymphocytes of leukemic patients as well as the susceptibility of their acute myeloid (AML) and lymphoblastic (ALL) leukemia cells to autologous and allogeneic LAKs were examined. In addition, neoplastic cells at diagnosis and at relapse were compared in the same patients for several features, including in vitro susceptibility to LAKs and to the drugs used in the induction phase, expression of MDR phenotype and of adhesion molecules, and differentiation markers. The NK activity of patients' LAK cells on K562 was significantly lower than that of a group of healthy donors whereas no differences were found in LAK activity as evaluated on Daudi cells. Three of 5 AML and 3 of 4 ALL were significantly more susceptible to autologous and allogeneic LAK lysis when blasts obtained at relapse were compared with leukemic cells of the same patients at diagnosis. This different lysability was not associated with in vitro modified sensitivity to drugs used in induction treatment. Moreover, no elevation in the expression of the multidrug-resistance (MDR)-related P170 glycoprotein was noted in relapsing leukemic cells. Even the expression of adhesion molecules and differentiation markers did not correlate with lysability of leukemic cells. These data demonstrate that relapsing leukemic blasts can be significantly lysed by LAK cells and suggest a rationale for adoptive immunotherapy with IL-2 and LAK cells in the treatment of acute leukemic patients.     

Impact of IL-2 and IL-2R SNPs on Proliferation and Tumorkilling Activity of Lymphokine-Activated Killer Cells from Healthy Chinese Blood Donors

One of the goals of tumor immunotherapy is to generate immune cells with potent anti-tumor activity throughin vitro techniques using peripheral blood collected from patients. However, cancer patients generally havepoor immunological function. Thus using patient T cells, which have reduced in vitro proliferative capabilitiesand less tumor cell killing activity to generate lymphokine-activated killer (LAK) cells, fails to achieve optimalclinical efficacy. Interleukin-2 (IL-2) is a potent activating cytokine for both T cells and natural killer cells.Thus, this study aimed to identify optimal donors for allogeneic LAK cell immunotherapy based on singlenucleotide polymorphisms (SNP) in the IL-2 and IL-2R genes. IL-2 and IL-2R SNPs were analyzed using HRMPCR.LAK cells were derived from peripheral blood mononuclear cells by culturing with IL-2. The frequencyand tumor-killing activity of LAK cells in each group were analyzed by flow cytometry and tumor cell killingassays, respectively. Regarding polymorphisms at IL-2-330 (rs2069762) T/G, LAK cells from GG donors hadsignificantly greater proliferation, tumor-killing activity, and IFN-γ production than LAK cells from TT donors(P<0.05). Regarding polymorphisms at IL-2R rs2104286 A/G, LAK cell proliferation and tumor cell killing weresignificantly greater in LAK cells from AA donors than GG donors (P<0.05). These data suggest that either IL-2-330(rs2069762)T/G GG donors or IL-2R rs2104286 A/G AA donors are excellent candidates for allogeneicLAK cell immunotherapy.     

Antitumor effect of large doses IL-2-activated HLA haploidentical peripheral blood stem cells on refractory metastatic solid tumor treatment

OBJECTIVE: The traditional immunotherapy for patients with refractory metastatic solid tumors is limited because tumors induce immunosuppression. New treatment is, therefore, needed. The aim of this study was to evaluate the clinical efficacy of infusion of high-dose interleukin (IL)-2-activated allogeneic haploidentical peripheral blood stem cells (haplo-PBSCTs) on patients with an advanced stage of refractory solid tumors. METHODS: This study involved 11 patients with refractory metastatic tumors and haploidentical relatives as donors for haplo-PBSCs. The therapeutic outcome of the IL-2-activated haplo-PBSC infusion and patients' cytokine levels were evaluated. The cytotoxicity of IL-2-activated haplo-PBSCs for tumor cells was determined using in vitro cytotoxicity assays. RESULTS: A range from 2.5 to 5.6 x 10(10) of activated haplo-PBSCs were harvested after exposure to rhIL-2, along with a significant increase in the proportion of natural killer (NK) cells and activated lymphocytes (CD69+ and CD25+), and enhanced cytotoxicity of haplo-PBSCs for several tumor cell lines. Following treatment, 1 (1/11) patient achieved a partial response (PR), 1 (1/11) achieved a mild response (MR), 6 (6/11) achieved stable disease (SD), and 3 (3/11) achieved progressive disease (PD). For all of the 11 patients, the median progression-free survival (PFS) was 5 months (3-14 months). We also observed the phenomenon of Th2 shifted to Th1, which played a crucial role in cancer immunotherapy. CONCLUSIONS: The adoptive transfusion of IL-2-activated haplo-PBSCs has potent antitumor effects both in vitro and in vivo. This finding suggests that IL-2-activated haplo-PBSCs may serve as an alternative therapy for advanced-stage solid tumors, especially for those patients who are refractory or ineligible for chemo- or radiotherapy.