Identification of Human Tumor Antigens Recognized by T-Cells and Their Use for Immunotherapy

Identification of human tumor antigens led to development of new immunotherapies for patients with cancer. Identification of T-cell epitopes allowed us to monitor antitumor T-cell responses quantitatively and qualitatively during immunotherapy as well as to develop more efficient immunotherapies with sufficient amounts of antigens in more immunogenic forms. Various immunotherapies, passive immunotherapies, including adoptive transfer of tumor reactive T-cells or allogeneic antigen-specific donor T-cells, and active immunization with identified tumor antigens or dendritic cells pulsed with tumor antigens are being evaluated in clinical trials. Although tumor regression has been observed in some patients, further improvement is required.     

Natural Killer Cell Alloreactivity in Haploidentical Hematopoietic Stem Cell Transplantation

Natural killer (NK) cells are primed to kill by several activating receptors. NK cell killing of autologous cells is prevented because NK cells coexpress inhibitory receptors (killer cell immunoglobulin-like receptors [KIR]) that recognize groups of (self) major histocompatibility complex class I alleles. Because KIRs are clonally distributed, the NK cell population in any individual are constituted of a repertoire with a variety of class I specificities. NK cells in the repertoire mediate alloreactions when the allogeneic targets do not express the class I alleles that block them. After haploidentical hematopoietic transplantation, NK cell-mediated donor-versus-recipient alloresponses reduce the risk of relapse in acute myeloid leukemia patients while improving engraftment and protecting against graft-versus-host disease. High-resolution molecular HLA typing of recipient and donor, positive identification of donor KIR genes, and, in some cases, functional assessment of donor NK clones identify haploidentical donors who are able to mount donor-versus-recipient NK alloreactions.     

急、慢性病毒性乙型肝炎病人干扰素-自然杀伤细胞系统功能的研究

为了研究干扰素自然杀伤细胞系统在病毒性肝炎发病机制中的作用，本文对４０例急性肝炎，２１例慢性肝炎和６例重症肝炎病人及１９名献血员外周血ＮＫ细胞活力和ＩＦＮ－γ 外诱生能力进行观察，结果表明急肝病人ＮＫ细胞活力与正常对照比较明显降低，慢肝病珍ＮＫ活力亦低下但无显著性差别两者ＩＦＮ－γ体外生能力与正常对照比较无差别，降低的ＮＫ细胞活力与ＩＦＮ－γ活力无相关关系。重症肝炎病人ＮＫ活 ＩＦＮ＿γ     

Graft versus Leukemia in 2023

Allogeneic hematopoietic stem cell transplantation (HSCT) is commonly utilized in the management of leukemia across multiple subtypes. Graft versus leukemia (GVL) is a critical component of successful transplantation and involves donor cells eradicating residual leukemia within the recipient. Graft versus host disease (GVHD) by contrast is a common complication of the transplantation process in which donor cells identify the recipient's various organ systems as foreign, thereby leading to a multitude of organ toxicities that can be described as autoimmune in nature. As both GVL and GVHD are mediated by a similar mechanism, these processes are felt to occur in tandem with one another. Here, we review the allogeneic HCT process in the context of GVL.     

Timing of Effective Natural Killer Cell Collection in Cancer Patients After High Dose Chemotherapy,Compared to Peripheral Blood Stem Cell Collection

Abstract: Natural killer (NK) and lymphokine-activated killer (LAK) activities in patients with seminoma were determined, and the timing of effective NK cell collection was evaluated, compared to autologus peripheral blood stem cell (PBSC) collection. After chemotherapy, patients were injected with granulocyte-colony stimulating factor (G-CSF). CD56 cells decreased in parallel with white blood cells and then were restored. After discontinuing the G-CSF, the CD56 cells increased and reached peak 3–5 days after PBSC collection. The peak showed higher levels than those at premedication. Recovery-peripheral blood lymphocytes (PBL) which were obtained 4 days after PBSC collection showed higher NK and LAK activities than the PBL obtained before chemotherapy (Before-PBL). They exhibited a high expression of CD2, CD 18, CD29, and CD49d. Furthermore, the LAK cells generated from the Recovery-PBL could aquire 9.6 folds of killer unit compared with the Before-PBL-LAK. These results suggest that, in order to obtain large numbers of NK cells, Recovery-PBL collection may be more advantageous than PBL collection in no relation with the chemotherapy.     

健康老年人外周血自然杀伤细胞活性的研究

本文应用~(125)IUdR标记K_(562)靶细胞技术测定健康老年人(60～87岁)和健康青年人(21～28岁)外周血淋巴细胞自然杀伤(NK)活性。结果表明,健康老年人NK细胞活性与健康青年人相比没有显著性差异(P>0.05)。对照前人的研究,该结果认为NK细胞活性不存在明显的随龄变化。     

A Novel Method to Quantify and Characterize Leukemia-Reactive Natural Killer Cells in Patients Undergoing Allogeneic Hematopoietic Stem Cell Transplantation following Conventional or Reduced-Dose Conditioning

The antitumor activity of natural killer (NK) cells has recently been shown to be assessable at a single-cell level via flow cytometric detection of CD107a. We used this novel method to prospectively quantify and characterize tumor-reactive NK cells in patients undergoing myeloablative or nonmyeloablative conditioning and allogeneic hematopoietic stem cell transplantation (HSCT). Degranulation of NK cells in the peripheral blood of 34 patients after HSCT (day +30, day +90) was determined by evaluating CD107a expression after coincubation of the cells with tumor targets. The percentage of degranulating NK cells was higher after nonmyeloablative conditioning than after myeloablative conditioning (P<.001), indicating a higher activation state and increased antitumor activity of NK cells after nonmyeloablative conditioning. We were able to analyze NK cell subsets separately and found that CD56bright NK cells following HSCT are functionally different from CD56bright NK cells in healthy donors, as indicated by a high percentage of degranulating NK cells in response to tumor targets in patients after HSCT. The CD107a assay is a new and feasible method to quantify and characterize tumor-reactive NK cells after HSCT. Using this method, we found that NK cells had high antitumor cytotoxic activity after nonmyeloablative conditioning, which may contribute to the effectiveness of nonmyeloablative conditioning.     

Omeprazole Inhibits Natural Killer Cell Functions

This study was designed to determine the possible effects of omeprazole on human natural killer cells. Peripheral venous blood samples were taken from 20 peptic ulcer patients before and at the 14th and the 28th days of omeprazole treatment. Mononuclear cells were removed from blood and their capability of making conjugation with K562 target cells and lysing K562 target cells was evaluated. A significant decrease was found (P < 0.001) in the 14th and the 28th days compared with the basal value of the capability of the mononuclear cells to conjugate with the K562 target cells and to lyse them. This study demonstrated that omeprazole significantly reduces natural killer cell functions. This finding suggests that omeprazole may also have some effects on the other systems in addition to parietal cell acid secretion.     

Melatonin-Induced Suppression of Human Lymphocyte Natural Killer Activity in Vitro

One of the important immune functions influenced by neuroendocrine factors is natural killer (NK) activity, which is directed against neoplastic and virus-infected cells. The effects of melatonin (Mel) and N-acetylserotonin (NAc-5HT) on NK activity of human peripheral blood lymphocytes were investigated. Leukocytes of healthy human subjects were used in the experiment. NK activity was estimated by measurement of radioactive chromium (51Cr) release from human leukemia cells K 562 (target cells). The previous exposure of human lymphocytes (effector cells) to Mel in concentrations of 10(-6) M and 10(-10) M resulted in an inhibition of NK activity (P less than 0.01) for all the examined effector-target cell ratios (10:1; 20:1, 40:1). NK activity was also suppressed by Mel (10(-8) M), but only if effector-target ratio equal to 20:1 was used (P less than 0.02), and by Mel (10(-12) M) for effector-target ratio equal to 40:1 (P less than 0.05). In none of the examined concentrations did NAc-5HT inhibit NK activity of human lymphocytes. On the basis of the data reported above, a direct suppressive effect of Mel (but not of NAc-5HT) on NK activity of human peripheral blood lymphocytes can be assumed.     

Natural killer cells – new understanding of basic biology may lead to more effective allogeneic haematopoietic stem cell transplantation

The natural killer (NK) cells are part of the innate immune system and are responsible for initial defences in the surveillance against malignant cells and virally infected cells. In addition to direct cytotoxicity, cytokines produced by NK cells amplify the immune response and help control the neoplasm/pathogen. Several activating and inhibitory receptors responsible for NK cell activation are recently characterized and play a crucial role in tumour eradication. These include, but are not limited to, the killer immunoglobulin-like receptors, C-type lectin receptors and natural cytotoxicity receptors. The downstream signalling of some of these receptors is also characterized. The net balance in the sum of the signals generated by ligation of activating and inhibitory receptors determines the final outcome, cytotoxicity versus tolerance. NK cell-based immunotherapy can be successfully exploited in the haematopoietic stem cell transplantation for the treatment of haematological malignancies and has a potential to separate the beneficial graft versus leukaemia effect from, often dangerous, graft versus host disease. This article reviews the NK receptors important in NK-mediated cytotoxicity in allogeneic haematopoietic stem cell transplantation.     

Aggressive Natural Killer Cell Leukemia: Report of a Chinese Series and Review of the Literature

Aggressive natural killer cell leukemia (ANKL) is a rare Epstein-Barr virus (EBV)-associated fulminating disease that is widely disseminated at diagnosis. Because of its typically extranodal presentation, differing degrees of NK cell involvement, and varying bone marrow pathology, ANKL can be confused with a reactive process. These features, coupled with a rapidly fatal course, have hampered systematic study of the pathogenesis of ANKL. Nine cases of ANKL were diagnosed and characterized by a single laboratory over a 2-year period. Constant features at presentation included disseminated disease, high fever, bone marrow involvement, and a high lactate dehydrogenase index. All cases were positive for EBV early region protein and negative for latent membrane protein 1, and all had a germline T-cell receptor gene configuration. Peripheral blood counts were variable, with severe thrombocytopenia being the most frequently encountered abnormality (7 of 9 cases). Hematophagocytosis, dyserythropoiesis, and stromal degeneration were the most frequent findings in the bone marrow. Neoplastic cells in the bone marrow were consistently CD2+, CD56+, CD45+, CD34-, CD117-, CD4-, and surface CD3-. Most cases were HLA-DR+ (8/9) and CD8- (8/9). Complex clonal cytogenetic abnormalities were found in 8 of 9 cases. Because of its aggressive course, rapid and accurate diagnosis of ANKL is essential for a better understanding of the etiology, pathogenesis, and treatment of the disease.     

类风湿关节炎患者外周血、滑液及滑膜组织中自然杀伤细胞的特征

目的探讨自然杀伤细胞（NK）在类风湿关节炎（RA）中的特征以及在疗效判断中的作用。方法采用流式细胞仪在单个细胞水平分析RA患者外周血、滑液与滑膜组织中NK细胞的数量。分析NK细胞中信号分子的改变以及产生细胞因子的能力，观察经肿瘤坏死因子（TNF）单克隆抗体治疗后RA患者外周血中NK细胞数量的变化与疗效之间的相关性。结果与健康对照者相比，RA患者外周血中CD3^-CD56^＋NK细胞的数量显著减少（8．95±3．72vs.5．31±5．07，P〈0．05）。RA患者滑液与滑膜组织中CD3^-CD56^＋、CD3^-CD247^＋NK细胞的数量均显著少于相应的外周血（P〈0．05），同时这些细胞中信号分子CD247表达的强度也显著降低。IL-12联合IL-18刺激使1．4％的RA外周血单个核细胞产生干扰素γ，其中1．1％为NK细胞产生；肉豆蔻酸酯和钙离子载体可以使15．33％的单个核细胞产生干扰素γ，其中13．87％为T淋巴细胞产生，NK细胞占1．47％。经TNF单克隆抗体治疗14周后，临床判定为良好反应和中度反应的5例患者中4例外周血中NK细胞数目增加，而治疗反应差的2例患者中NK细胞数量增加或减少各1例。结论RA患者循环与炎症部位中NK细胞的数量显著减少，这些细胞的信号传导和细胞因子产生能力低下；NK细胞的低下状态与所处环境中的炎症程度有关，TNF单克隆抗体治疗在改善患者疾病活动性的同时，使外周血中NK细胞呈增加趋势，NK细胞可能成为疗效判断的一项指标。     

Role of Natural Killer and Gamma-Delta T cells in West Nile Virus Infection

Natural Killer (NK) cells and Gamma-delta T cells are both innate lymphocytes that respond rapidly and non-specifically to viral infection and other pathogens. They are also known to form a unique link between innate and adaptive immunity. Although they have similar immune features and effector functions, accumulating evidence in mice and humans suggest these two cell types have distinct roles in the control of infection by West Nile virus (WNV), a re-emerging pathogen that has caused fatal encephalitis in North America over the past decade. This review will discuss recent studies on these two cell types in protective immunity and viral pathogenesis during WNV infection.     

干扰素α-2b联合利巴韦林治疗慢性丙型肝炎患者肝内免疫细胞的变化

目的观察干扰素α-2b和利巴韦林联合治疗前后漫性丙型肝炎患者肝内免疫细胞动态变化情况，为研究免疫调节疗法提供依据。方法利用流式细胞计数仪对42例慢性丙型肝炎患者应用联合治疗前及其中11例治疗后患者末梢血和（或）肝脏组织进行分析。结果与对照组相比慢性丙型肝炎组肝脏内CD56^＋、CD57^＋、CD161^＋细胞及CD56^＋T淋巴细胞阳性率明显减少（P〈0．01），CD161^＋T淋巴细胞有减少倾向；CD56^＋T淋巴细胞表达的CD28可见减少，CD152的表达可见增加（P〈0．05）；CD83^＋CD1a^＋细胞阳性率有减少倾向，CD80^＋CD11c^＋、CD86^＋CD11c^＋细胞阳性率明显减少（P〈0．01）；显效组减低的CD56^＋、CD161^＋、CD56T、CD161^＋T、CD80^＋CD11c^＋、CD86^＋CD11c^＋细胞治疗后可见增加。结论慢性丙型肝炎患者肝脏内自然杀伤细胞、自然杀伤T淋巴细胞数量及树突状细胞数量和功能减低，联合治疗使细胞免疫功能的抑制得到了改善。     

Which factors influence the development of GVHD in HLA-matched or mismatched transplants?

The sheer diversity of HLA alleles makes the probability of finding matched unrelated donors for patients requiring hematopoietic cell transplantation (HCT) a complex situation. New evidence suggests that mismatching at certain HLA loci may provide a greater benefit in terms of graft-versus-leukemia effect than other mismatches when HLA-matched donors are not available. This review summarizes the current understanding of HLA matching requirements for unrelated donor HCT.     

Natural killer cell activities of patients with breast cancer against different target cells

Summary Natural killer (NK) cell activity against K 562 erythroleukemic- and MCF-7 breast carcinoma-derived cells was monitored in short-term (3h/ K 562) and long-term (18h/MCF-7) chromium release tests for 60 patients with untreated primary breast disease. Target cell lysis was the same for patient groups with benign (n=13) and malignant (n=47) breast disease (27% versus 36% mean chromium release; target: effector ratio 40:1 for K 562 and 28% versus 40% for MCF-7 cells). NK activity as defined by short-term lysis of K 562 cells did not correlate with MCF-7 cell lysis in long-term assays for the carcinoma patients. This functional heterogeneity of natural cytotoxic activities of breast cancer patients was confirmed by a different age distribution for K 562 and MCF-7 cell lysis and high levels of MCF-7-directed NK activity in the grade I tumor group (56.2%). Our results indicate that measurement of peripheral blood NK activity against a breast carcinoma-derived cell line (MCF-7) defines a disease-related natural cytotoxic acitivity which correlates better with prognostic tumor parameters (tumor grading) than NK activity as defined by the lysis of K 562 erythroleukemic cells. NK activity testing against breast carcinoma cell lines should be used to monitor natural cytotoxic activities in breast cancer patients and its modulation by different routes of treatment.Supported by Jubiläumsfond der österreichischen Nationalbank, Project No. 2227     

Role of NK,NKT cells and macrophages in liver transplantation

Liver transplantation has become the treatment of choice for acute or chronic liver disease. Because the liver acts as an innate immunity-dominant organ, there are immunological differences between the liver and other organs. The specific features of hepatic natural killer(NK), NKT and Kupffer cells and their role in the mechanism of liver transplant rejection, tolerance and hepatic ischemia-reperfusion injury are discussed in this review.     

Therapeutic Effect of Repeated Natural Killer T Cell Stimulation in Mouse Cholangitis Complicated by Colitis

Primary sclerosing cholangitis is often complicated by ulcerative colitis. Recently, we reported on Th1-dominant cholangitis associated with experimental colitis, and natural killer T (NKT) cells might play an important role in this model. The aim of this study was to clarify the immunopathogenic role of NKT cells in this model using α-galactosylceramide. CD-1 mice were administered 2.0% dextran sulfate sodium for 29 days and injection of α-galactosylceramide was performed every 5 days, then inflammation was assessed. Mononuclear cells from the liver were analyzed with respect to cytokine production and the surface marker. α -Galactosylceramide improved survival rate, weight gain, and inflammation score. Also, interferon-γ release from MNC, CD4/CD8 ratio, NKT cell population, and NK cell population were decreased by this treatment. These findings indicate that repeated stimulation of NKT cells modifies the Th1/Th2 balance to reduce Th1 dominance, and this may be a mechanism by which α -galactosylceramide has a therapeutic effect.     

Roles of liver innate immune cells in nonalcoholic fatty liver disease

Nonalcoholic fatty liver disease (NAFLD) has become the most common liver disease in the United States and other developed countries and is expected to increase in the next few years. Emerging data suggest that some patients with NAFLD may progress to nonalcoholic steatohepatitis (NASH), cirrhosis and even hepatocellular carcinoma. NAFLD can also promote the development and progression of disease in other organ systems, such as the cardiovascular and endocrine (i.e. diabetes) systems. Thus, understanding th...     

Immunotherapy of human leukemia with antibody to pluripotential K-562 stem cells

Summary Gamma () globulin was fractionated from the serum of a goat immunized with the pluripotential leukemia cell line K-562. The lyophilized -globulin preparation, termed leukoglobulin, contained about 50% immune IgG and suppressed the proliferation of heterotransplanted leukemia and lymphoma cells of human origin. The main aims of this study were to evaluate the potential therapeutic value of leukoglobulin and to determine its toxicity in humans with terminal leukemia and patients whose disease was unresponsive to current therapy. Two patients with CML, one with AMML, four with All, and one with AML were treated once a week for up to 5 weeks with leukoglobulin intravenously at doses ranging from 2 to 29 mg/kg. Leukoglobulin was well tolerated with minimal adverse effects and produced an initial mobilization of blasts from the bone marrow, spleen, and other organs with a parallel increase in the number of blasts in the systemic circulation. Subsequent injections of leukoglobulin led to a sharp decrease and the eventual eradication of blasts from the peripheral blood and bone marrow. Except in patients with CML, immature cells other than blasts also markedly diminished. The results of the clinical trials indicated a synergism with or potentiation of most chemotherapeutic agents used. Two possible uses for a combination of leukoglobulin and antileukemic drugs are indicated by the results were report here; drug-antibody synergism for cases showing no response to chemotherapy alone or leukoglobulin given immediately after chemotherapy is administered to eliminate residual leukemia cells. Alternatively, leukoglobulin can be given as a single therapeutic agent during the induction or maintenance phases of treatment to patients with leukemia resistant to other therapeutic combinations.Abbreviations CML chronic myelogenous leukemia - AMML acute myelomonocytic leukemia - ALL acute lymphoblastic leukemia - AML acute myclogenous leukemia - LAA leukemia-associated antigens - IgG, IgA, IgM, IgD immunoglobulins G, A, M, and D - i.v. intravenous - BUN blood urea nitrogen - BUS busulfan - 6-MP 6-mercaptopurine - VCR vincristine - DNM daunomycin - PRED prednisone - MTX methotrexate (amethopterin) - CPM cyclophosphamide - CIC cerebral intermittent claudication - PBS phosphate-buffered saline These clinical trials were supported in part by the Fundación Roux-Ocefa, Buenos Aires, Argentina. The basic research was supported by Grants 18185 and 17533 awarded by the National Cancer Institute, DHHS, USA. The clinical trials have been approved by the Argentina Public Health Service (No. 22020000052098778/3) and by the University of Tennessee (CRP No. 878 and IRB No. 1445) National Cancer Institute Number 678, DHHS, USA Comp. of Tumor Immunotherapy Protocols 7:77, 1979