Factors associated with successful mobilization of progenitor hematopoietic stem cells among patients with lymphoid malignancies

PurposeThis study's main aim was to assess the effect of 2 mobilization regimens (granulocyte colony-stimulating factor [G-CSF] and chemotherapy vs. G-CSF alone) on the yield of CD34⁺ cells in the apheresis components of patients with lymphoid malignancy. We also sought to identify possible predictors of CD34⁺ cell yield in the apheresis components.Patients and MethodsCD34⁺ cells were mobilized and harvested from 89 patients with non-Hodgkin lymphoma (n = 62) or Hodgkin disease (n = 27). Forty-one patients (46.1%) were mobilized with G-CSF, and 48 (53.9%) were mobilized with chemotherapy and G-CSF. Univariate and multivariate analyses were used to examine potential predictors of the CD34⁺ cell yield (collection of > 2.7 > 10⁶ cells/kg), such as the number of peripheral CD34⁺ cells, age, sex, diagnosis, disease stage, weight, bone marrow status at baseline, mononuclear cells, white blood cells, and platelet counts.ResultsThe median patient age was 41 years (range, 12-66 years), and the median patient weight was 72 kg (range, 44-123 kg). Mobilization of peripheral blood progenitor cells (PBPCs) was superior when using chemotherapy and G-CSF versus G-CSF alone (3.6 > 10⁶ cells/kg vs. 2.2 > 10⁶ cells/kg; P = .001). CD34⁺ cell counts and platelet counts in the peripheral blood significantly correlated with CD34⁺ yield (P < .01 and P = .009, respectively). The yield was also significantly affected by weight, diagnosis, mobilization regimen, and baseline bone marrow status (P = .021, P = .05, P = .002, and P = .043, respectively).ConclusionMany factors influence harvesting of PBPCs, including diagnosis, bone marrow status at baseline, patient weight, and the type of mobilization regimen. The number of CD34⁺ cells in the peripheral blood can be used to predict the timing of apheresis and optimize yield.     

NK1.1+ Cells and CD8+ T Cells Mediate the Antitumor Activity of Cl-IB-MECA in a Mouse Melanoma Model

Cl-IB-MECA, synthetic A₃ adenosine receptor agonist, is a potential anticancer agent. In this study, we have examined the effect of Cl-IB-MECA in a mouse melanoma model. Cl-IB-MECA significantly inhibited tumor growth in immune-competent mice. Notably, the number of tumor-infiltrating NK1.1⁺ cells and CD8⁺ T cells was significantly increased in Cl-IB-MECA-treated mice. This effect was correlated with high levels of tumor necrosis factor α (TNF-α) and interferon γ in melanoma tissue. Depletion of either CD8⁺ T cells or NK1.1⁺ cells completely abrogated the antitumor effect of Cl-IB-MECA. Accordingly, Cl-IB-MECA did not affect tumor growth in nude mice. In addition, we also found that the number of mature and active conventional dendritic cells at the tumor site was increased after Cl-IB-MECA administration. Moreover, Cl-IB-MECA significantly increased TNF-α and IL-12p40 release from splenic CD11c⁺ cells. In conclusion, our study provides novel insights into the mechanism by which Cl-IB-MECA leads to an effective antitumor response that involves the activation of natural killer cells and CD8⁺ T cells and further highlights its therapeutic potential.     

宫颈癌患者外周血T淋巴细胞亚群和NK细胞数量与临床分期的相关性分析

目的 探讨宫颈癌患者外周血T淋巴细胞亚群和NK细胞数量的变化及其与宫颈癌分期的相关性.方法 收集2010年6月-2011年6月期间在沈阳医学院奉天医院妇科治疗的146例宫颈癌患者及同期在我院进行体检的50例健康女性(对照组),宫颈癌患者根据临床分期分为0期、Ⅰ期、Ⅱ期、Ⅲ期、Ⅳ期5组.采用免疫荧光法和流式细胞仪测定各组患者外周血T淋巴细胞亚群和NK细胞数量.结果 宫颈癌患者CD3+,CD3+ CD4+,NK细胞数量及CD3+ CD4+/CD3+ CD8+比值均低于对照组,且随分期增加而逐渐降低;而CD3+ CD8+细胞数量高于对照组,且随分期增加而逐渐增高.CD3+、CD3+CD4+、NK细胞数量及CD3+ CD4+/CD3+ CD8+细胞比值与宫颈癌临床分期呈负相关(P＜0.01);CD3+ CD8+细胞数量与宫颈癌分期呈正相关(P＜0.01).结论 宫颈癌患者细胞免疫功能低下,且临床分期越晚,免疫功能越低,检测T淋巴细胞亚群、NK细胞可用于宫颈癌患者的免疫监测.     

Chimeric antigen receptor‐engineered cytokine‐induced killer cells overcome treatment resistance of pre‐B‐cell acute lymphoblastic leukemia and enhance survival

Pre‐emptive cancer immunotherapy by donor lymphocyte infusion (DLI) using cytokine‐induced killer (CIK) cells may be beneficial to prevent relapse with a reduced risk of causing graft‐versus‐host‐disease. CIK cells are a heterogeneous effector cell population including T cells (CD3⁺ CD56^?), natural killer (NK) cells (CD3^?CD56⁺) and natural killer T (T‐NK) cells (CD3⁺ CD56⁺) that exhibit non‐major histocompatibility complex (MHC)‐restricted cytotoxicity and are generated by ex vivo expansion of peripheral blood mononuclear cells in the presence of interferon (IFN)‐γ, anti‐CD3 antibody, interleukin‐2 (IL‐2) and interleukin‐15 (IL‐15). To facilitate selective target‐cell recognition and enhance specific cytotoxicity against B‐cell acute lymphoblastic leukemia (B‐ALL), we transduced CIK cells with a lentiviral vector encoding a chimeric antigen receptor (CAR) that carries a composite CD28‐CD3ζ domain for signaling and a CD19‐specific scFv antibody fragment for cell binding (CAR 63.28.z). In vitro analysis revealed high and specific cell killing activity of CD19‐targeted CIK/63.28.z cells against otherwise CIK‐resistant cancer cell lines and primary B‐ALL blasts, which was dependent on CD19 expression and CAR signaling. In a xenograft model in immunodeficient mice, treatment with CIK/63.28.z cells in contrast to therapy with unmodified CIK cells resulted in complete and durable molecular remissions of established primary pre‐B‐ALL. Our results demonstrate potent antileukemic activity of CAR‐engineered CIK cells in vitro and in vivo, and suggest this strategy as a promising approach for adoptive immunotherapy of refractory pre‐B‐ALL.     

Mobilization of peripheral blood stem cells using regimen combining docetaxel with granulocyte colony-stimulating factor in breast cancer patients

Objective:To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor(G-CSF) in breast cancer patients.Methods:A total of 57 breast cancer patients were treated with docetaxel 120 mg/m2.When the white blood cell(WBC) count decreased to 1.0×109/L,patients were given G-CSF 5-g/kg daily by subcutaneous injection until the end of apheresis.Peripheral blood mononuclear cells(MNC) were isolated by Cobe Spectra Apheresis System.The percentage of CD34+ cell was assayed by flow cytometry.Results:At a median 6 of days(range 3-8) after the administration of docetaxel,the median WBC count decreased to 1.08×109/L(range 0.20-2.31).The median duration of G-CSF mobilization was 3 days(range 2-7).The MNC collection was conducted 8-12 days(median 10 days) after docetaxel treatment.The median MNC was 5.35×108/kg(range 0.59-14.07),the median CD34+ cell count was 2.43×106/kg(range 0.16-16.69).The CD34+ cell count was higher than 1.00×106/kg in 47 of 57 cases(82.46%) and higher than 2.00×106/kg in 36 cases(63.16%).The CD34+ cell count was higher than 2.00×106/kg in 27 collections(23.68%).The MNC count and the CD34+ cell count were correlated with the bottom of WBC after docetaxel chemotherapy(r=0.364,0.502,P=0.005,0.000).The CD34+ cell count was correlated with the MNC count(r=0.597,P=0.000).The mobilization and apheresis were well tolerated in all patients.Mild perioral numbness and numbness of hand or feet were observed in 3 cases.No serious adverse events were reported.Conclusion:Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.     

Lipoteichoic acid of Bifidobacterium in combination with 5-fluorouracil inhibit tumor growth and relieve the immunosuppression

To investigate the therapeutic efficacy of lipoteichoic acid of Bifidobacterium (BLTA) in combination with 5-fluorouracil (5-FU) treatment on the mice bearing inoculated hepatoma-22 (H₂₂) cells and the effects of BLTA on immunological regulation of organism, and explore its mechanisms.MethodsTumor-bearing mice were treated with 5-FU alone, BLTA alone or BLTA in combination with 5-FU. The tumor size were observed and measured regularly. The growth inhibiting rate (IR) of tumor was detected. MTT assay was used to evaluate the proliferation of T lymphocytes and splenic NK cell and CTL activity. Enzyme linked immunosorbent assay (ELISA) was used to detect the change of IFN-Γ. FCM was used to detect T subgroup ratio of spleen cells of tumor-bearing mice. Expression change of mRNA and proteins of Foxp3 and TIM-3 were detected by Real-Time-PCR and Western blot in tumor-bearing mice tumor tissue.ResultsBoth 5-FU and BLTA had inhibition effect on tumor-growth. While in the 5-FU + BLTA group, the inhibition of tumor growth was more significant, with increased T lymphocyte proliferation and IFN-Γproduction of spleen cells. Spleen cells of tumor-bearing mice had high CD4⁺CD25⁺regulatory T cell (CD4⁺CD25⁺T_reg) ratio and high mRNA and proteins expression of Foxp3 and TIM-3, but in the BLTA and 5-FU group, CD4⁺CD25⁺T_reg ratio degraded, with down regulation mRNA and proteins expression of Foxp3 and TIM-3. But CD4⁺ T cells also decreased in spleen cells of tumor-bearing mice by alone 5-FU treated, splenic NK cell and CTL activity also degraded, while CD4⁺ T cells and splenic NK cell and CTL activity significantly increased by BLTA treated. BLTA in combination with 5-FU could also enhance the ratio of CD4⁺ T cells and splenic NK cell and CTL activity.ConclusionThe present study suggested that BLTA in combination with 5-FU could enhance antitumor effect, with inhibiting TIM-3/TIM-3L pathway, cutting down immunosuppressive activity of CD4⁺CD25⁺ T_reg and enhancing cell-mediated immunity.     

The Proangiogenic Phenotype of Natural Killer Cells in Patients with Non-Small Cell Lung Cancer

The tumor microenvironment can polarize innate immune cells to a proangiogenic phenotype. Decidual natural killer (dNK) cells show an angiogenic phenotype, yet the role for NK innate lymphoid cells in tumor angiogenesis remains to be defined. We investigated NK cells from patients with surgically resected non-small cell lung cancer (NSCLC) and controls using flow cytometric and functional analyses. The CD56⁺CD16^- NK subset in NSCLC patients, which represents the predominant NK subset in tumors and a minor subset in adjacent lung and peripheral blood, was associated with vascular endothelial growth factor (VEGF), placental growth factor (PIGF), and interleukin-8 (IL-8)/CXCL8 production. Peripheral blood CD56⁺CD16^- NK cells from patients with the squamous cell carcinoma (SCC) subtype showed higher VEGF and PlGF production compared to those from patients with adenocarcinoma (AdC) and controls. Higher IL-8 production was found for both SCC and AdC compared to controls. Supernatants derived from NSCLC CD56⁺CD16^- NK cells induced endothelial cell chemotaxis and formation of capillary-like structures in vitro, particularly evident in SCC patients and absent from controls. Finally, exposure to transforming growth factor-β₁ (TGFβ₁), a cytokine associated with dNK polarization, upregulated VEGF and PlGF in peripheral blood CD56⁺CD16^- NK cells from healthy subjects. Our data suggest that NK cells in NSCLC act as proangiogenic cells, particularly evident for SCC and in part mediated by TGFβ₁.     

A Phase I Study of the Safety and Feasibility of Bortezomib in Combination With G-CSF for Stem Cell Mobilization in Patients With Multiple Myeloma

BackgroundWe previously reported that administration of bortezomib (BTZ) after 4 days of granulocyte colony-stimulating factor (G-CSF) significantly augments mobilization in mice. We hypothesized that administration of BTZ at peak G-CSF mobilization in patients with multiple myeloma (MM) would be safe, augment mobilization, and have an in vivo purging effect on circulating myeloma cells.Patients and MethodsThis was a phase I study using 3 dose levels of BTZ. G-CSF was administered for 5 days. On the evening of the fourth day, a single dose of BTZ was administered. Peripheral blood was drawn 1 to 2 hours before and 15 to 18 hours after BTZ administration (before day 5 G-CSF administration) to analyze the mobilization effect of BTZ. Standard apheresis was then performed starting on day 5. After mobilization, patients underwent autologous stem cell transplantation (ASCT) per institutional guidelines.ResultsTen patients were enrolled. There were no dose-limiting toxicities. Median peripheral blood CD34⁺ cells at day 4 before BTZ administration was 16 per microliter and 15 hours later was 32 per microliter suggesting that administration of BTZ at peak G-CSF mobilization augments the mobilization effect of G-CSF. The effect of BTZ on circulating MM cells was unclear. All patients had successful engraftment after ASCT.ConclusionAdministration of 1 dose of BTZ at peak G-CSF mobilization was safe and well tolerated, enhanced stem cell mobilization, and did not affect graft viability. The mobilization effect of BTZ at peak G-CSF mobilization shown in this phase I study needs to be confirmed in a larger randomized trial.     

Numbers and cytotoxicities of CD3+CD56+ T lymphocytes in peripheral blood of patients with acute myeloid leukemia and acute lymphocytic leukemia

Recent reports have highlighted the role of cellular immunity in anti-tumor defenses. T lymphocytes are known to play important part in anti-cancer immunity. The number and function of T lymphocytes are altered in chronic leukemia patients. CD3⁺CD56⁺ T lymphocytes have also been found to be abnormal in cancer patients. We therefore investigated changes in the number and cytotoxicity of CD3⁺CD56⁺ T lymphocytes in the peripheral blood of acute leukemia (AL) patients (excluding acute promyelocytic leukemia), to improve our understanding of the role of this T lymphocyte subset. We analyzed CD3⁺CD56⁺ T lymphocyte numbers and cytotoxicities in healthy controls, AL patients, and AL patients with complete remission. Lymphocyte counts were performed in peripheral blood and flow cytometry was used to determine cell numbers and cytotoxicities. The absolute number of CD3⁺CD56⁺ T lymphocytes was increased in AL patients (including acute myeloid [AML] and acute lymphocytic leukemia [ALL]) compared with healthy controls (P < 0.05), but their functioning was significantly reduced (P < 0.05). The number of CD3⁺CD56⁺ T lymphocytes in AML and ALL patients who achieved remission following chemotherapy was close to healthy controls (P > 0.05), but their functioning was still significantly reduced (P < 0.05). In addition, the number of CD3⁺CD56⁺ T lymphocytes increased significantly in AML patients with increased peripheral blood white blood cell (WBC) counts, and in ALL patients without increased WBCs. These results suggest that cellular immunity may respond to AML and ALL, but that lymphocyte cytotoxicity remains impaired. Dysfunction of CD3⁺CD56⁺ T lymphocytes in AML and ALL patients may contribute to the failure of the host immune response against leukemic blasts.     

Low miR-16 expression induces regulatory CD4+NKG2D+ T cells involved in colorectal cancer progression

MiR-15a/16 is a member of the miRNA cluster that exhibits tumor suppression and immune modulation via targeting multiple genes. Decreased miR-15a/16 expression is involved in many cancer cells. Here, miR-16 had decreased expression in NK1.1^-CD4⁺NKG2D⁺ T cells and bound with the 3’-UTR of NKG2D gene. MiR-15a/16-deficient mice had many CD4⁺NKG2D⁺ T cells, which produced TGF-β1 and IL-10 and inhibited the IFN-γ production of CD8⁺ T cells. Adoptive transfer of NK1.1^-CD4⁺NKG2D⁺ T cells from miR-15a/16-deficient mice promoted tumor growth in vivo. However, no changes for NK1.1^-CD4⁺NKG2D⁺ T cells were found in the miR-15a/16-transgenic mice. Although the miR-15a/16 transgenic mice transplanted with B16BL6 or MC38 cells exhibited rapid growth, these tumor-bearing mice did not show changes in NK1.1^-CD4⁺NKG2D⁺ T cell distributions in either spleens or tumors. When NK1.1^-CD4⁺ T cells were stimulated by α-CD3/sRAE-1 ex vivo, the NKG2D expression was difficult to induce in the T cells of miR-15a/16-transgenic mice. Finally, increased frequencies of regulatory CD4⁺NKG2D⁺ T cells with low miR-16 levels were observed in patients with late-stage colorectal cancer (Duke’s C, D). Thus, miR-16 modulates NK1.1^-CD4⁺NKG2D⁺ T cell functions via targeting NKG2D. Low miR-16 expression in CD4⁺ T cells induces the regulatory CD4⁺NKG2D⁺ T subpopulation, which promotes tumor evasion via the secretion of immune-suppressive molecules.     

CD4+ T cells play a crucial role for lenalidomide in vivo anti-tumor activity in murine multiple myeloma

Lenalidomide modulates the host immune response against myeloma via multiple actions. Although these effects have been elucidated in vitro, the central action of lenalidomide-mediated anti-myeloma immune response in vivo is not clear. To investigate its immune action in vivo, we selected the murine myeloma cell line 5TGM1, which is resistant to direct tumoricidal effects of lenalidomide in vitro and in immunodeficient mice, but sensitive to lenalidomide treatment in 5TGM1-bearing immunocompetent mice. Depletion of CD4⁺ T cells, but not NK cells, B cells, or CD8⁺ T cells, deprived lenalidomide of its therapeutic effects on 5TGM1-bearing immunocompetent mice. Lenalidomide significantly increased the numbers of IFN-γ-secreting CD4⁺ and CD8⁺ T cells but had no effects on NK cells and B cells in this mouse model. Lenalidomide slightly decreased the number of CD25⁺Foxp3⁺ T cells but increased perforin expression in CD8⁺ T cells in vivo. Using this mouse model for investigation of anti-tumor immune action of lenalidomide, we demonstrated that lenalidomide facilitated a type-1 anti-tumor immune response in vivo. The CD4⁺ T cell subset may play a critical role in the lenalidomide-mediated anti-myeloma immune response in vivo.     

Anti-G-CSF treatment induces protective tumor immunity in mouse colon cancer by promoting NK cell,macrophage and T cell responses

Granulocyte colony-stimulating factor (G-CSF) is a cytokine that is highly expressed in human and mouse colorectal cancers (CRC). We previously reported that G-CSF stimulated human CRC cell growth and migration, therefore in this study we sought to examine the therapeutic potential of anti-G-CSF treatment for CRC. G-CSF is known to mobilize neutrophils, however its impact on other immune cells has not been well examined. Here, we investigated the effects of therapeutic anti-G-CSF treatment on CRC growth and anti-tumor immune responses. C57BL/6 mice treated with azoxymethane/dextran sodium sulfate (AOM/DSS) to induce neoplasms were administered anti-G-CSF or isotype control antibodies three times a week for three weeks. Animals treated with anti-G-CSF antibodies had a marked decrease in neoplasm number and size compared to the isotype control group. Colon neutrophil and macrophage frequency were unchanged, but the number of macrophages producing IL-10 were decreased while IL-12 producing macrophages were increased. NK cells were substantially increased in colons of anti-G-CSF treated mice, along with IFNγ producing CD4⁺ and CD8⁺ T cells. These studies are the first to indicate a crucial role for G-CSF inhibition in promoting protective anti-tumor immunity, and suggest that anti-G-CSF treatment is a potential therapeutic approach for CRC.     

Changes in peripheral lymphocyte subsets in patients after partial microwave ablation of the spleen for secondary splenomegaly and hypersplenism: A preliminary study

Purpose: Microwave ablation therapy for secondary splenomegaly and hypersplenism has been shown to be effective from pre-clinical animal models and clinical investigations. This study was performed to determine its effects on the status of peripheral lymphocyte subsets in patients receiving microwave ablation of the spleen.

Materials and methods: Ten patients with secondary splenomegaly and hypersplenism received microwave ablation of the spleen during laparoscopy or percutaneously under ultrasound guidance. The percentage peripheral blood T cells, B lymphocytes and NK cells were measured using flow cytometry before and on days 1, 3 and 7 after therapy, as well as 1 and 3 months afterwards.

Results: Percentages of CD3⁺ and CD4⁺ cells increased rapidly 1 month after therapy. There was no significant change in CD8⁺, CD4⁺/CD8⁺ or NK cells of the pre- and post-therapy levels and B lymphocytes increased significantly after therapy. In patients with an ablation volume (AV) less than 20% (group A), T cells increased 1 month after ablation but decreased 3 months after ablation. B lymphocytes increased significantly after surgery. Levels of NK cells were lower than that before therapy on each testing. In patients with 20–40% AV (group B), levels of T cells, B lymphocytes and NK cells showed an increase. Levels of CD4⁺ cells were significantly higher in group B than in group A, 3 months after therapy.

Conclusions: Microwave ablation therapy for splenomegaly and hypersplenism appears to have a favourable effect on peripheral lymphocyte subsets. A relationship may exist between the ablation volume and the level of peripheral lymphocyte subsets.     

Transgenic expression of IL-33 activates CD8 T cells and NK cells and inhibits tumor growth and metastasis in mice

IL-33 is a multifunctional cytokine in immune regulation that activates Th1 cells, Th2 cells, CD8⁺ T cells and NK cells. Our study showed that transgenic expression of IL-33 attenuated tumor metastasis in the B16 melanoma and Lewis lung carcinoma (LLC) metastatic models. The percentages and cytotoxicity of CD8⁺ T cells and NK cells and their infiltration into the tumor tissues were significantly increased by the transgenic expression of IL-33 in tumor-bearing mice. Treatment with recombinant IL-33 could also increase the cytotoxicity of CD8⁺ T cells and NK cells in vitro. In addition, depletion of CD8⁺ T cells and NK cells using anti-CD8 or anti-asialo GM1 antibody abolished the pulmonary metastasis inhibition mediated by IL-33. Furthermore, IL-33 stimulated the activation of NF-κB and increased CD69 expression, which is a marker of the activated form of the two cell subsets, in CD8⁺ T cells and NK cells. Our results suggest that IL-33 stimulated NF-κB signaling and promoted the proliferation, activation and infiltration of CD8⁺ T cells and NK cells, which resulted in the inhibition of pulmonary metastasis in B16 melanoma and LLC mice models.     

Pilot trial of interleukin-2 with granulocyte colony-stimulating factor for the mobilization of progenitor cells in advanced breast cancer patients undergoing high-dose chemotherapy: expansion of immune effectors within the stem-cell graft and post-stem-cell infusion.

PURPOSE: To evaluate whether administration of interleukin-2 (IL-2) with granulocyte colony-stimulating factor (G-CSF) improves mobilization of immune effector cells into the stem-cell graft of patients undergoing high-dose chemotherapy and autografting. PATIENTS AND METHODS: We performed a trial of stem-cell mobilization with IL-2 and G-CSF in advanced breast cancer patients receiving high-dose chemotherapy with cyclophosphamide, thiotepa, and carboplatin and stem cells followed by IL-2. The trial defined immune, hematologic, and clinical effects of IL-2 in this setting. RESULTS: Of 32 patients enrolled, nine received G-CSF alone for mobilization. Twenty-one of 23 patients mobilized with IL-2 plus G-CSF had stem cells collected with more mononuclear cells than those receiving G-CSF (19.3 v 10.4 x 10(8)/kg; P =.006), but fewer CD34(+) progenitor cells (6.9 v 22.0 x 10(6)/kg; P =.049). The IL-2 plus G-CSF-mobilized patients had greater numbers of activated T (CD3(+)/CD25(+)) cells (P =.009), natural killer (NK; CD56(+)) cells (P =.007), and activated NK (CD56 bright(+)) cells (P: =.039) than those patients mobilized with G-CSF. NK (P =.042) and lymphokine-activated killer (LAK) (P =.016) activity was increased in those mobilized with IL-2 + G-CSF, whereas G-CSF-mobilized patients had a decline in cytolytic activity. In the third week posttransplantation, immune reconstitution was superior in those mobilized with IL-2 plus G-CSF based on greater numbers of activated T cells (P =.003), activated NK cells (P =.04), and greater LAK activity (P =.003). The 16 of 21 IL-2 + G-CSF-mobilized patients with adequate numbers of stem cells (> 1.5 x 10(6) CD34(+) cells/kg) collected engrafted rapidly posttransplantation. CONCLUSION: The results demonstrate that G-CSF + IL-2 can enhance the number and function of antitumor effector cells in a mobilized autograft without impairing the hematologic engraftment, provided that CD34 cell counts are more than 1.5 x 10(6) cells/kg. Mobilization of CD34(+) stem cells does seem to be adversely affected. In those mobilized with IL-2 and G-CSF, post-stem-cell immune reconstitution of antitumor immune effector cells was enhanced.     

A distinct distribution of natural killer cell subgroups in human tissues and blood

Natural killer (NK) cells have been subdivided according to their CD16/ CD56 expression into at least 2 subgroups. We examined the distribution of these NK subgroups in humans. In the blood of normal individuals, CD16⁺⁺/ CD56⁺/ CD3^– NK cells predominate, constituting more than 90% of all NK cells. In contrast, decidua is infiltrated almost exclusively by CD16^+/–/ CD56⁺/ CD3^– NK cells (>90%), a fact so far seen in context with decidua being an immunoprivileged tissue. However, this NK subgroup can also be detected in the blood, where it comprises about 10% of NK cells. We have found that normal (colon) as well as neoplastic (ovarian and urothelial carcinoma) tissues are also predominantly infiltrated by this CD16^+/– NK subgroup. Lymphatic fluid draining solid tissues contains CD16^+/– NK cells exclusively, with absolute numbers of NK cells being very low. No predominating NK subgroup was seen in ascites. CD16^+/– NK cells, when tested against the target cell lines K562 and JAR, revealed a cytotoxic spectrum different from CD16⁺⁺ NK cells and from T cells. A change in the CD16/ CD56/ CD3 phenotype was not seen in either subgroup in long-term cultures containing IL-2 (1,000 U/ ml). Our data indicate that the decidua is not the only solid tissue infiltrated by CD16^+/– NK cells. Other normal and malignant tissues were also infiltrated predominantly by this NK cell subgroup. We suggest that CD16^+/– NK cells represent a functionally distinct NK subgroup involved in the surveillance of solid tissues. Int. J. Cancer 78:533–538, 1998. © 1998 Wiley-Liss, Inc.     

Prognostic significance of CD169‐positive lymph node sinus macrophages in patients with endometrial carcinoma

Lymph node (LN) macrophages play critical roles in anti‐tumor immunity, which develops via the activation of cytotoxic T cells (CTL) and NK cells. The present study aims to determine the prognostic significance of CD169⁺ LN macrophages in patients with endometrial carcinoma (EC). The number of CD169⁺ cells or the CD169⁺‐to‐CD68⁺ macrophage ratio in regional LN (RLN), and the number of CD8⁺ CTL or CD57⁺ NK cells in tumor tissues were investigated by immunohistochemistry in paraffin‐embedded tissue samples from 79 patients with EC. A high density of CD169⁺ cells in the RLN of patients with EC was correlated with an early clinical stage or no LN metastasis. A high number of CD169⁺ cells and a high CD169⁺‐to‐CD68⁺ macrophage ratio were significantly associated with longer overall survival in EC. We also found that the density of CD169⁺ macrophages was positively correlated with the number of CD8⁺ CTL and CD57⁺ NK cells that infiltrated into tumor tissues. A high density of CD57⁺ cells in EC tissues was associated with a better prognosis, while a high density of CD8⁺ cells was not linked to an altered prognosis. The present study showed that the density of CD169⁺ macrophages in RLN was associated with an improved prognosis in EC patients. CD169⁺ macrophages in RLN might represent a useful marker for assessing clinical prognoses and monitoring anti‐tumor immunity in patients with EC.     

脐血体外扩增干祖细胞和同时诱导分化T、NK和DCs免疫细胞

Umbilical cord blood stem cell transplantation (CB- SCT) has approached significant success in treatment of lethal congenital or malignant disorders, but CBSCT with low incidence of GVHD is associated with higher rates of delayed or failed engraftment and relapse than bone marrow transplants. This may be caused by immature im- mune cells, compared with the corresponding cells from adults. More studies have been reported that cord blood T lymphocytes are immature in phenotype and function…     

Prediction of tumor recurrence by distinct immunoprofiles in liver transplant patients based on mass cytometry

Recurrence of hepatocellular carcinoma (HCC) after liver transplantation (LT) is a marker of poor prognosis. However, the reliable biomarkers of post-LT HCC recurrence remain to be identified. In this study, serial peripheral blood samples from the LT recipients with and without HCC recurrence were collected at five time points. Single-cell mass cytomertry (CyTOF) was utilized for the in-depth analysis of peripheral blood monocellular cells (PBMCs). CyTOF analysis showed that at 3 weeks post-LT, the activated immune cell population was increased, while the fraction of immune cells with suppressive functions (myeloid-derived suppressive cells) was reduced. The post-LT immune composition in patients with LT for HCC was enormously different from that in patients with LT for causes other than HCC. Furthermore, at 3 weeks after LT, compared with patients without recurrence, the patients with HCC recurrences were high in two subsets of T cells: CD57⁺ HLA-DR⁺ CD8⁺ and CD28⁺γδ. The CD57⁺ HLA-DR⁺ CD8⁺ T cells presented high levels of perforin, granzyme B, and Ki-67 and displayed a highly cytotoxic and proliferative phenotype, while the CD28⁺γδ T cells had reduced levels of IFN-γ and, hence, were less activated compared to CD28^- cells. Based on these findings, we concluded that analyzing the PBMCs of LT recipients by CyTOF can predict the post-LT HCC recurrence. The distinct immune features can stratify patients with the risk of HCC recurrence at 3 weeks after LT, which will help clinician in further management plan and improve the prognosis of patients.