rIL2／LAK细胞净化自体骨髓移植诱生皮肤移植物抗宿主病一例

ｒＩＬ２／ＬＡＫ细胞净化自体骨髓移植诱生皮肤移植物抗宿主病一例刘启发，刘晓力，周淑芸，伍柏松，陈琪自体骨髓移植（ＡＢＭＴ）治疗白血病主要不足是复发率高，其原因与微小残留白血病（ＭＲＤ）和缺乏移植抗白血病（ＧｖＬ）效应有关。我们对４例行ＡＢＭＴ的白血病...     

净化及非净化自体骨髓移植治疗白血病39例临床研究

报告自体骨髓移植治疗白血病３９例，其中非净化自体骨髓移植（ＡＢＭＴ）１４例，净化自体骨髓移植（ＰＡＢＭＴ）２５例。中位年龄２８岁（１０～４３岁）。ＡＭＬ２７例，ＡＬＬ１０例，ＣＭＬ２例。ＣＲ１３１例，ＣＲ２７例，ＮＲ１例。ＣＲ至移植时间中位数６．７个月（２～１９个月）。预处理方案：ＴＢＩ加Ａｒａ－ｃ、ＤＮＲ或ＶＰ１６。ＡＢＭＴ组及ＰＡＢＭＴ组３年无病生存（ＤＦＳ）率分别为６８．３２％及６７．５７％，复发率为３０．７６％及２６．８０％。但ＰＡＢＭＴ组ＡＭＬ患者３年ＤＦＳ率为８２．３５％及ＣＲ２期移７５％３年ＤＦＳ率为７５％，明显高于ＣＲ。期移植未净化者５０％。化疗组３年ＤＦＳ率为７．３８％及复发率７６．４％，两移植组疗效优于化疗组。     

单克隆抗体体外大容量骨髓净化白血病的研究

应用补体介导细胞毒法体外消除２例白血病骨髓中残存瘤细胞，并作自身骨髓移植。用国产ＣＤ７单抗（ＬＮ６７－８及ＬＳ９３－２，ＩｇＧ２ｂ）首次净化１例Ｔ－ＡＬＬ骨髓及ＣＤ１０单抗（５５，ＩｇＧ２ａ）净化１例ＣＡＬＬＡ＋－ＡＬＬ。靶细胞清除率均达３个对数级，骨髓ＣＦＵ－ＧＭ回收率分别为５７．６％～７５．８％及５１．７％～７２．３％。Ｔ－ＡＬＬ病人移植后２６天获造血重建。现已完全缓解３年，首次证实国产单抗ＬＮ６７－８及ＬＳ９３－２净化骨髓有效、安全。     

异基因和同基因骨髓移植治疗55~66岁的骨髓增生异常综合征病人

作者用骨髓移植(ＢＭＴ)治疗５０例骨髓增生异常综合征(ＭＤＳ)病人,年龄５５．３~６６．２(中位５８．８)岁。按照ＦＡＢ标准,１３例难治性贫血(ＲＡ),１９例ＲＡ伴原始细胞增多(ＲＡＥＢ),１６例ＲＡＥＢ转化型或急性髓系白血病(ＲＡＥＢ－ｔ/ＡＭＬ),２例慢性粒单细胞白血病(ＣＭＭＬ)。按     

血液病患者外周血IL－2活性的观察

采用ＣｏｎＡ诱导的外周血淋巴细胞产生的白细胞介素Ⅲ（ＩＬ－２），用活化的Ｃ５７ＢＬ／ｂ小鼠脾细胞为反应细胞，以３Ｈ－ＴｄＲＤＮＡ掺入法测定５５例血液病患者ＩＬ－２活性，结果表明：①白血病及淋巴瘤、多发性骨髓瘤和由实体瘤引起的继发性贫血３３例其外周血ＩＬ－２活性与正常对照组相比明显降低（Ｐ＜０．００１）。②原发性血小板减少性紫癜（ＩＴＰ）７例其ＩＬ－２活性与对照组差异显著（Ｐ＜０．００１），其Ｔ细胞亚群ＯＫＴ４／Ｔ８＜Ｉ和Ｌ－２活性降低呈正相关，ＯＫＴ８百分值明显高于正常。     

同种LAK细胞、5－FU等抗癌剂腹腔注入治疗原发性肝癌腹水的临床观察

本文采用同种ＬＡＫ细胞联合ＩＬ－２腹腔注入（ＩＰ），５－ＦＵ联合ＭＭＣ、ＣＢＤＣＡ腹腔注入，同种ＬＡＫ细胞联合ＩＬ－２静脉输入以及放腹水、利尿、补充白蛋白等方法，分别治疗晚期ＰＨＣ腹水患者共４０例，结果表明，同种ＬＡＫ细胞联合ＩＬ－２经ＩＰ（１０例），５－ＦＵ联合ＭＭＣ、ＣＢＤＣＡ经ＩＰ（１０例），均使４例（４０％）腹水减少或消失、症状明显改善；同种ＬＡＫ细胞联合ＩＬ－２全身治疗（１０例），以及放腹水、利尿、补充白蛋白等一般治疗（１０例），均仅使１例（１０％）腹水减少，疗效欠佳；结果提示，腹腔内ＬＡＫ联合ＩＬ－２治疗及腹腔内５－ＦＵ联合ＭＭＣ、ＣＢＤＣＡ化疗，是值得推荐的治疗晚期肝癌腹水的手段之一。     

CD3AK细胞联合IL—2脂质体抗鼠肝转移瘤的实验研究

目的 观察ＩＬ－２脂质体对ＣＤ３ＡＫ体内抗肿瘤的增强作用。方法：利用Ｇ５７ＢＬ／６小鼠脾细胞与抗ＣＤ３单克隆抗体和白细胞介素－－－２（ＩＬ－２）共同培养制备ＣＤ３ＡＫ细胞（ＣＤ３ＭｃＡｂ ａｃｔｉｖａｔｅｃｄ ｋｉｌｌｅｒ ｃｅｌｌｓ）；从脾内注射Ｂ１６黑色素瘤细胞制备鼠肝转移瘤模型：用ＰＣ和ＣＨ合成ＩＬ－２复合脂质体；ＣＤ３ＡＫ细胞联合ＩＬ－２脂质体用于体内抗Ｂ１６黑色素瘤肝转移。结果：共同培养     

异基因外周血造血干细胞移植治疗白血病

我院从１９９６年４月至１９９８年７月,用异基因外周血干细胞移植（ＡｌｌｏＰＢＳＣＴ）成功地治疗了１５例白血病患者。一、资料与方法１－病例：１５例患者中急性髓细胞白血病（ＡＭＬ）８例,急性淋巴细胞白血病（ＡＬＬ）５例,慢性粒细胞白血病（ＣＭＬ）慢性期２例;急性白血病１２例处在完全缓解期（ＣＲ）,１例为第３次复发。男１１例,女４例,年龄１４～４９岁,中位数３９岁。２－供体：１４例系患者的同胞,ＨＬＡＡ、Ｂ及ＤＲ配型完全相合１３例,ＨＬＡ－Ａ位点不合１例,母亲供体１例,ＨＬＡＢ位点不相合。ＡＢＯ…     

用RT—PCR和Genescan分析CML急变期人TCRVβT细胞的表达和克隆性

目的：了解ＣＭＬ急变期的ＴＣＲＶβ亚家族Ｔ细胞的表达及其克隆性。方法：采用ＲＴ－ＰＣＲ扩增４例ＣＭＬ急变期病人的外周血单个核细胞的ＴＣＲＶβ２４个亚家族的互补决定区３（ＣＤＲ３），产物进一步经基因扫描分析确定Ｔ细胞的克隆性，结果：病人外周血仅表达４－９个Ｖβ亚家族Ｔ细胞，主要为Ｖβ１，Ｖβ２，Ｖβ３，Ｖβ５和Ｖβ１７；基因扫描分析显示２例ＣＭＬ急粒变的部分产物为寡克隆性，而２例ＣＭＬ急粒变的部分产     

联合应用rIL—2和LAK细胞治疗小鼠H22肝癌和B16黑色素瘤…

本研究通过对立小鼠Ｈ２２肝癌和Ｂ１６黑色素瘤肺转移瘤模型，并联合应用ｒＩＬ－２和ＬＡＫ细胞对该两种肺转移瘤模型进行治疗，以观察ＬＡＫ细胞在体内抗肿瘤转移瘤的作用。单用ｒＩＬ－２和ＬＡＫ细胞＋ｒＩＬ－２治疗组均较对照组明显降低瘤结节转移数（Ｐ〈０．００１），又以联合应用ＬＡＫ细胞加ｒＩＬ－２治疗效果最佳。     

Lymphokine-activated killer (LAK) cell purging of leukemic bone marrow: range of activity against different hematopoietic neoplasms

Natural killer cells activated in vitro by incubation with IL-2 display a broad range of cytolytic activity against neoplastic cells. These lymphokine-activated killer (LAK) cells can discriminate between neoplastic and normal bone marrow cells and may represent a useful means of purging bone marrow prior to autologous transplantation. We demonstrate that LAK cells can successfully remove four distinctly different malignant hematopoietic cell types from normal bone marrow grafts. The LAK purging technique is capable of a 2-3 log10 reduction in tumor cells in the bone marrow graft without compromising hematological recovery or survival. Our results also suggest, however, that an inhibitory effect on stem cell function by allogeneic LAK cells exists, and this form of purging may be used only if greater levels of bone marrow are transferred in an allogeneic setting. The ability to detect and eliminate malignant cells in bone marrow prior to use for autologous transplantation suggests that LAK cells, alone or in conjunction with current methods of bone marrow purging, could be useful for the in vitro treatment of bone marrow in patients who require high-dose chemotherapy and autologous bone marrow transplantation.     

Autologous transplantation of bone marrow purged in vitro with anti-CD7- (WT1-) ricin A immunotoxin in T-cell lymphoblastic leukemia and lymphoma

Seven patients with high-risk acute T-cell lymphoblastic leukemia (T- ALL) and six with T cell lymphoma (T-LL) were treated with autologous bone marrow transplantation (ABMT) after in vitro purging of their bone marrow with WT1 (CD7)-ricin A-chain immunotoxin. CD7 expression on the tumor cells showed large variations between the individual patients and was highly related to the specific cytotoxicity of WT1-ricin A. Incubation of bone marrow with up to 10(-8)mol/L WT1-ricin A in the presence of 6 mmol/L NH4Cl did not compromise the growth potential of the hematopoietic progenitors CFU-GM, CFU-GEMM, and BFU-E. Hematologic engraftment (greater than 10(9) leukocytes/L) occurred within a normal time period (median, 17 days). Seven patients are alive and in complete remission (CR) at 48+, 44+, 40+, 26+, 11+, 7+, and 6+ months after ABMT. Four patients relapsed within 6 months after ABMT. Two of them had the lowest CD7 expression on their tumor cells, the other two were transplanted in CR2 and CR3. Two patients died from transplantation related infections. The immunologic reconstitution was delayed, although the numbers of T cells reached normal levels within 1 month. The number of CD7+ cells remained low up to 1 year after transplantation. The T4/T8-ratio was decreased for at least 6 months. The T-cell response to mitogens recovered to normal levels after 1 year. This study shows that ABMT with WT1-ricin A purged bone marrow in high-risk T-cell malignancies results in a complete hematopoietic and a delayed immunologic reconstitution. The actuarial relapse free survival is 61% at 3 years.     

Monoclonal antibody purging and autologous bone marrow transplantation in acute myelogenous leukemia in complete remission

A mouse monoclonal antibody (S4-7) reacting with human myelomonocytic cells has been previously shown to be suitable for bone marrow purging in selected acute myelogenous leukemia (AML) patients with S4-7 positive leukemic clonogenic cells at diagnosis. The results obtained in seven AML patients who underwent such a treatment, followed by autologous bone marrow transplantation (ABMT), are now reported. Six patients underwent ABMT in first complete remission (CR), one in second CR, after BAVC conditioning regimen. One patient died of infection 1 month after ABMT; in the other six a complete recovery of hemopoiesis was observed. In spite of S4-7 reactivity with normal myelomonocytic cells, a prompt recovery of granulopoiesis was however observed both in in vitro liquid culture and in vivo with a median time of 20 days to reach granulocyte values of 500 x 10(6)/l. The patient transplanted in 2nd CR relapsed 3 months after ABMT. Of the five evaluable patients transplanted in 1st CR, two relapsed 8 and 9 months post-ABMT while three remain in continuous CR at 35, 47, 57 months. Leukemic cells of two of the three patients with recurrent disease were studied at relapse and in both could be detected a significant percentage of S4-7 negative cells, detectable neither at diagnosis nor (one patient) at the time of first relapse after standard chemotherapy.     

Interleukin 2 induction of lymphokine-activated killer (LAK) activity in the peripheral blood and bone marrow of acute leukemia patients. I. Feasibility of LAK generation in adult patients with active disease and in remission

The feasibility of in vitro interleukin 2 (IL-2) activation and expansion of mononuclear cells (MNCs) derived from adult patients with acute myelogenous leukemia (ANLL) was studied. Patients' natural killer (NK) and lymphokine-activated killer (LAK) cell activity was compared with that of normal donors in terms of: (a) cytolytic activity (four- hour 51Cr release assay) against an NK-sensitive target (K562), NK- resistant targets (Raji/Daudi), and fresh/cryopreserved autologous and allogeneic leukemic blasts; (b) proliferation and expansion in culture with 1,000 U/mL recombinant IL 2 (rIL 2); and (c) the cell surface phenotype of the cultured cells. In 21 of 24 patients with active disease (AP) MNCs derived from the peripheral blood (PBL) or bone marrow (BM) could be cultured and expanded in the presence of rIL 2. These cultures initially contained between 30% and 50% blasts, and during 2 to 4 weeks of culture destruction of blasts and enrichment of up to 60% in cells with the morphology of large granular lymphocytes (LGLs) was observed. Expansion in culture varied between two- and 100- fold. MNCs from all patients in remission (RP) could be activated by rIL 2 and expanded up to 30-fold after 1 to 3 weeks in culture. NK activity of fresh PBLs from AP was significantly lower than in normal controls, whereas NK activity of RP was within the normal range. High levels of postactivation NK and LAK activity on K562/Raji/Daudi and on fresh/cryopreserved leukemic blasts was generated in approximately 50% of cases of AP and in most RP. Cell surface phenotype studies showed that cultured cells derived from ANLL patients were significantly enriched (up to 40%) in NKH-1 (Leu 19) positive cells, with RP LAK cells also expressing a high proportion of CD16 positive cells (up to 40%). This study has shown that it is feasible to activate and significantly expand killer cells derived from active disease and remission ANLL patients during 1 to 3 weeks culture with IL 2 with good maintenance of cytolytic activity. Both initial NK activity and LAK generation was optimal in remission patients. Based on data from this study, a clinical protocol has been developed for treatment of early relapse ANLL patients with LAK cells cultured for 1 to 3 weeks and systemic IL 2.     

Intensive cytotoxic therapy followed by autologous bone marrow transplantation for non-Hodgkin's lymphoma of high-grade malignancy

Fourteen patients with non-Hodgkin's lymphoma (NHL) of high-grade malignancy were treated with cyclophosphamide and total body irradiation followed by autologous bone marrow transplantation (ABMT). All patients were pretreated with conventional chemotherapy. Three of four patients with drug-resistant disease achieved complete remission (CR), but relapse occurred within six months. Four patients in partial remission (PR) achieved CR; one died because of sepsis, two relapsed within six months, and one is still in CR 28+ months later. Six were treated in CR, five in first CR, and one in second CR. From these six patients (who received this treatment as consolidation therapy), five are in unmaintained CR seven to 31+ months after ABMT (one patient died of a secondary illness). There were two therapy-related deaths, both in patients with a poor clinical condition. Toxicity of this treatment was mild for those receiving transplants who were in better condition. These preliminary results suggest that intensive cytoreductive therapy followed by ABMT may improve disease-free survival in patients in NHL of high-grade malignancy in CR.     

Long-term follow-up of autologous bone marrow transplantation in patients with relapsed follicular lymphoma.

We report the results of high-dose chemoradiotherapy and anti-B-cell monoclonal antibody-purged autologous bone marrow transplantation (ABMT) in patients with relapsed indolent follicular lymphoma. Between March 1985 and May 1995, 153 patients underwent ABMT using a uniform ablative regimen with cyclophosphamide and total body irradiation and bone marrow (BM) purging. All patients received multiple chemotherapy regimens before ABMT. At BM harvest, only 30% of patients were in complete remission, and overt BM infiltration was present in 47%. The disease-free survival (DFS) and overall survival (OS) are estimated to be 42% and 66% at 8 years, respectively. Patients whose BM was negative by polymerase chain reaction (PCR) for bcl2/IgH rearrangement after purging experienced longer freedom from recurrence than those whose BM remained PCR positive (P <.0001). Continued PCR negativity in follow-up BM samples was also strongly predictive of continued complete remission (CR). The 12-year survival from diagnosis for these 153 patients is 69%. Considering that the median survival from diagnosis and first recurrence of patients with advanced follicular lymphoma are 8 and 5 years, respectively, our results provide evidence that myeloablative therapy and ABMT may prolong overall survival.     

Evaluation of cytoreductive therapy prior to high dose treatment with autologous bone marrow transplantation in relapsed and refractory Hodgkin's disease

Based on observations that bulky disease at autologous bone marrow transplantation (ABMT) may be correlated with poor outcome in Hodgkin's disease, we have assessed the ability of conventional-dose chemoradiotherapy to reduce tumour burden to a minimum prior to ABMT. Thirty-seven patients with relapsed or refractory Hodgkin's disease referred for intensive therapy and ABMT were treated initially with one to five cycles of DHAP chemotherapy. All patients had previously received MOPP and ABVD chemotherapy or similar regimens. Four patients achieved complete remission (CR) and 12 partial remission (PR), for a total response rate of 43%. Eight partial responders and four non-responders to DHAP achieved significant further tumour reduction with local radiotherapy (five CR, seven PR). Six of 10 non-responders to DHAP responded to alternative salvage chemotherapy (mini-BEAM, CEP or augmented CVP). Overall, 24/37 patients (65%) achieved effective cytoreduction (nine CR, 15 PR with minimal disease) and have proceeded to ABMT. Patients with bulky disease at relapse or limited stage (II, IIIA) at diagnosis were less likely to respond to DHAP, but some of these could be cytoreduced with alternative therapy. In addition, the number of prior chemotherapy regimens correlated inversely with likelihood of response to DHAP. The results indicate that approximately two-thirds of patients with Hodgkin's disease who relapse after MOPP and ABVD-like regimens can achieve effective cytoreduction with conventional-dose chemoradiotherapy and proceed to ABMT in CR or PR with minimal disease.     

Evaluation of lymphocyte subsets after autologous bone marrow transplantation with marrow treated by ASTA Z 7557 in acute leukemia: incidence of the in vitro treatment

The lymphocyte subset reconstitution after high-dose chemotherapy and total body irradiation followed by autologous bone marrow transplantation (ABMT) has been studied in ten patients with acute leukemia (AL) (6 ALL and 4 ANLL) in complete remission (CR). Bone marrow was treated in vitro with high-dose ASTA Z 7557, individually determined according to CFU-GM sensitivity. The different peripheral blood lymphocyte subsets were characterized by means of monoclonal antibodies (indirect immunofluorescence assay) belonging to the following classes of differentiation: OKT11-T11 (CD2), OKT3-T3 (CD3), OKT4-T4 (CD4), OKT8-T8 (CD8), OKIal-I2 (HLA-DR), Leu7 (natural killer/killer) and by means of polyspecific antiimmunoglobulin sera (direct immunofluorescence assay). Data in these ten patients were compared with those of a control group of 21 normal donors and with a control group of 14 patients in CR without ABMT. Our results showed a marked depression of the T4:T8 ratio in patients with AL before ABMT, compared with normal donors who had respective values of 1.02 and 1.33 (p less than 0.01). This depression was increased and prolonged up to day 515 after ABMT, with a value of 0.32 (p less than 0.01 compared with the pregraft situation; p less than 0.001 compared with normal donors). This T4:T8 ratio imbalance was related to the depletion of the T4+ population and to the increase of the T8+ subset. This imbalance was emphasized after ABMT. The Leu 7+ population was also increased in grafted patients compared with normal donors (p less than 0.01). The B-cell population remained unchanged throughout the study. We conclude that patients autografted with marrow treated in vitro by high-dose ASTA Z 7557 may experience a long-term T-cell subset imbalance.     

自体骨髓移植治疗成人恶性淋巴瘤后的长期随访

1983年至1991年我们用高剂量长春新碱、阿糖胞苷、环已亚硝脲及环磷酰胺加全淋巴(身)照射[Hd-VCCA+TL(B)I]和自体骨髓移植(ABMT)治疗了21例中高危组成人恶性淋巴瘤患者。其中5例为晚期,6例为耐药复发,4例为部分缓解(PR),4例为首次缓解(CR_1),2例为第二次缓解(CR_2)。平均随访37个月,9年生存率在何杰金氏淋巴瘤(HL)和非何杰金氏淋巴瘤(NHL)分别预期为89%和64%,提示该方案毒性反应可以耐受。如果在患者病程早期进行,可使约70%的成人恶性淋巴瘤患者长期存活。同时提示对骨髓受累或原淋巴细胞型的患者在移植前应作适当的净化残留肿瘤细胞的处理。     

Successful autologous bone marrow transplantation in second remission of malignant histiocytosis.

This report describes an 18-year-old man with disseminated malignant histiocytosis (MH). The patient initially attained complete remission (CR1) with conventional chemotherapy and then relapsed 14 months later. In second complete remission (CR2) 2 years and 8 months after initial diagnosis, an autologous bone marrow transplantation (ABMT) was undertaken following conditioning with the BEAM regimen. Bone marrow collected in CR2 was incubated with mafosfamide at a dose adjusted to the individual sensitivity of normal CFU-GM according to our current protocol. At the time of writing, 4 years post-transplant, this patient remains disease free. This is the first report of ABMT with marrow treated in vitro by mafosfamide in MH.