大鼠外周血自然杀伤细胞对肝细胞的细胞毒效应及大黄？…

运用３Ｈ－ＴｄＲ肝细胞内掺入法对大鼠外周血自然杀伤对肝细胞的细胞毒作用进行观察，证实自然杀伤细胞对肝细胞具有细胞毒作用（Ｐ〈０．００１）。大黄在１００μｇ／ｍｌ浓度水平能抑制此自然杀伤细胞的细胞毒作用（Ｐ〈０．０５）。     

膀胱癌外周血自然杀伤细胞活性的探讨

本文对9例膀胱癌患者采外周血自然杀伤细胞活性进行测定,同时与正常对照组比较,结果表明,病人组NK细胞活性明显降低(P<0.01)。鉴于干扰素和卡介苗对治疗膀胱癌均有疗效,可能是增强和改善了膀胱癌患者的NK细胞活性功能,而对治疗起到了积极作用。     

依托咪酯对人外周血单个核细胞凋亡及细胞毒效应的影响

目的探讨依托咪酯在体外对健康人外周血单个核细胞凋亡及细胞毒作用的影响。方法抽取健康人外周静脉血,梯度密度离心法分离采集单个核细胞体外培养,分为依托咪酯0.5μg/ml(E0.5组)、5μg/ml(E5组)、50μg/ml(E50组)和空白对照(C组)组,孵育24h后,以Annexin-V/碘化丙啶(PI)荧光探针标记流式细胞术分析各组细胞凋亡情况,并以人红白血病细胞系K562瘤细胞标准株为靶细胞,应用四甲基偶氮唑盐(MTT)比色法检测各组细胞毒效应的大小。结果与C组比较,E0.5、E5组单个核细胞凋亡率差异无统计学意义,而E50组凋亡显著增加(P<0.01)。E0.5、E5组单个核细胞对K562细胞毒作用差异亦无统计学意义,而E50组显著降低(P<0.05)。结论在临床有效血药浓度下,依托咪酯在体外环境下不影响单个核细胞的凋亡和对K562的细胞毒效应。     

巴比妥类麻醉下低温对大鼠自然杀伤细胞活性及肿瘤转移易感性的影响

目的　探讨低温对大鼠自然杀伤 (NK)细胞活性及肿瘤肺转移的影响。方法　雄性Wistar大鼠 3 0只随机分为低温 (3 2℃ )组、常温 (3 7.5℃ ,大鼠的正常体温 )组、对照组。检测不同温度组自然杀伤细胞活性 (NKCA)及肺转移瘤的数量。结果　低温组 (NK)细胞的活性明显低于常温组和对照组 (P <0 .0 5 )。低温组肺转移瘤的数量明显多于对照组 (P <0 .0 5 ) ,常温组与对照组相比无显著性差异。结论　巴比妥类麻醉下低温抑制了自然杀伤细胞的活性并增加了肺转移瘤的易感性。     

静脉注射免疫球蛋白治疗对复发性流产患者的外周血自然杀伤细胞及妊娠结局影响的Meta分析

目的 采用Meta方法来分析静脉注射免疫球蛋白(IVIG)在治疗复发性流产(RSA)对外周血自然杀伤(NK)细胞及妊娠结局的影响。方法 计算机检索PubMed、Web of Science、Embase、Cochrane Library和中国知网(CNKI)数据库,以复发性流产、反复流产、习惯性流产、静脉注射免疫球蛋白、自然杀伤细胞等为关键词进行文献检索,检索时间自建库至2022年4月。由2名评价员分别按照Cochrane手册、纽卡斯尔-渥太华量表(NOS)、Minors量表评价随机对照试验(RCT)、队列研究、无对照组的单臂试验研究的文献质量和偏倚风险;采用Rev Man5.4.1软件进行Meta分析。结果 本Meta分析共纳入14篇文献,包含1 393例患者。纳入的14篇文献中8篇为中质量文献,6篇为高质量文献。Meta分析结果显示,在改善外周血NK细胞数量方面,当一次性给药(1～5 d)时,IVIG治疗后NK细胞数量显著低于治疗前[I²=22%,95%CI(1.99,3.82),P<0.05];当连续给药时,无论是治疗至孕12周左右[I²     

腹腔感染大鼠肺泡巨噬细胞对肺细胞的细胞毒作用及下法对其影响

目的：为观察腹腔感染后肺泡巨噬细胞对肺细胞的胞毒作用及下法对其影响。方法：采用经ＬＰＳ活化的腹膜炎大鼠肺泡巨噬细胞与经３ＨＴｄＲ掺入的肺泡细胞及含大承气汤有效成分的血清进行细胞培养。结果：腹腔感染大鼠肺泡巨噬细胞对肺细胞的胞毒作用较对照组明显增强（Ｐ＜００５），经下法治疗４８ｈ后胞毒作用减弱（Ｐ＜００５），ＬＰＳ诱生大鼠肺泡巨噬细胞对肺细胞的胞毒作用明显增强（Ｐ＜００１），大承气汤能降低腹腔感染及经ＬＰＳ活化之大鼠肺泡巨噬细胞对肺细胞的胞毒作用（Ｐ＜００５）。结论：巨噬细胞在感染性炎症及脏器损伤过程中发挥着重要作用。大承气汤能减弱肺泡巨噬细胞活化后对肺泡上皮细胞的胞毒作用。     

红细胞生成素对血透患者自然杀伤细胞活性的影响

目的 探讨血透患者自然杀伤细胞(NK)活性与贫血之间的关系。方法 采用LDH释放法,对12例血透接受重组人红细胞生成素(rHuEPO)治疗前后血红蛋白(Hb)水平和NK活性进行观察。结果 经rHuEPO治疗后,患者Hb和NK活性均有明显提高,但rHuEPO对NK细胞并无刺激作用,而红细胞对NK细胞活性有明显的增强作用。结论 血透患者的NK细胞活性低下与贫血有关,rHuEPO提高NK细胞活性的作用是通过改善贫血而非rHuEPO的直接作用。     

酸性微环境下大蒜素增强大鼠自然杀伤细胞的功能活性

目的 观察体外酸性培养环境下大蒜素对自然杀伤(NK)细胞功能活性的影响,并探讨其机制.方法 以pH 5.6、pH 6.5和pH 7.2的完全RPMI 1640培养基对大鼠脾脏CD3-NKR-P1+NK细胞进行悬浮培养,并给予30 mg/L浓度的大蒜素进行处理.酶联免疫吸附试验(ELISA)检测细胞培养液中干扰素(IFN)-γ的分泌水平,流式细胞仪检测NK细胞的增殖和凋亡率,乳酸脱氢酶法检测NK细胞的功能活性.结果 酸性培养环境下大鼠脾脏NK细胞的增殖能力明显下降,NK细胞功能活性明显受阻.在pH 7.2、pH 6.5和pH 5.6三种不同的培养条件下,大蒜素对NK细胞增殖率的影响表现为随培养环境的pH降低反而升高,以pH 5.6、培养16 h时达最高33.3%.与之相对应,NK细胞分泌的IFN-γ达(64.59±0.09)ng/L,较培养4 h时升高28%,且对小鼠淋巴瘤Yac-1细胞的杀伤活性也达到最高水平.结论 大蒜素可通过提高NK细胞IFN-γ分泌水平明显改善酸性培养环境下NK细胞的功能活性.

Abstract：

Objective To investigate the role of allicin on rat nature killer (NK) cell activity in vitro under acidic microenvironment, and its possible mechanism. Methods CD3- NKR-P1+ NK cells isolated from the rat spleen were cultured in the complete RPMI 1640 medium ( pH 5. 6, pH 6. 5, or pH 7. 2 respectively), and treated with allicin at final concentration of 30 mg/L. Enzyme linked immunosorbent assay (ELISA) was used to determine supernatant interferon (IFN)-γ levels. The percentage of NK cells proliferation and apopotosis was analyzed by flow cytometry. NK cell cytotoxicity toward YAC-1 tumor cells was detected by LDH release assay. Results Proliferation and cytotoxicity of NK cells were significantly suppressed by acidic microenvironment in vitro. Under the cultured condition of acidic pH below 7. 2, allicin seemed to promote NK cells proliferation, which reached to highest level of 33% at pH 5. 6 cultured for 16 h. Correspondingly, at pH of 5. 6, allicin induced a marked increase of IFN-γ concentration in the supernatant from (50. 07 ± 0. 38) (cultured for 4 h) to (64. 59 ± 0. 09) ng/L ( cultured for 16 h). The cytotoxicity of NK cells toward YAC-1 tumor cells was also found strongest under the condition of pH 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK cells under the acidic cultured condition, which might be related to the increase of IFN-γ production.     

大黄对急性胰腺炎大鼠早期的治疗作用

采用胰管内逆行注入牛磺胆酸钠造成大鼠急性胰腺炎模型，造模后用中药大黄、生理盐水进行治疗，并观察治疗前后急性胰腺炎大鼠血中肿瘤坏死因子、内毒素及淀粉酶的变化，结果显示大黄组造模后２４小时后内毒素水平明显低于生理盐水组，大黄组肿瘤坏死因子、淀粉酶水平均较生理盐水组有显著降低。实验显示大黄对急性胰腺炎大鼠早期病生理改变有明显的治疗作用     

Natural Killer Cells and the Immune Response in Solid Organ Transplantation

Natural killer (NK) cells have been characterized classically for their cytotoxicity against pathogen infected or stressed cells as well as for their role in monitoring the expression of self MHC I. However, the participation of NK cells in solid organ transplantation (SOT) is poorly defined due to conflicting clinical and animal model data. Preclinical models have shown that NK cells exacerbate T‐cell allogeneic responses during rejection, but can also promote tolerance induction under immunosuppressive conditions. Further, while protocols such as costimulatory blockade effectively induce tolerance by blocking T‐cell activation and promoting Treg generation, how such regimens regulate other innate and adaptive immune cells, including NK cells, is incomplete. This review examines NK cells and the regulation of their effector functions in SOT.     

吸入麻醉对T细胞亚群和NK细胞的影响

观察异氟醚和安氟醚吸入麻醉对病人Ｔ细胞亚群、自然杀伤细胞的影响。方法：３８例行肺叶切除术的病人,随机分为异氟醚麻醉组２０例、安氟醚麻醉组１８例。术前、麻醉后１小时,术后１天、术后１周,术后２周分别检测血清Ｔ细胞亚群及ＮＫ细胞活性。结论安氟醚吸入麻醉对病人细胞免疫的抑制作用较异氟醚吸入麻醉明显。     

Activation of Natural Killer Cells and Macrophages by Porcine Endothelial Cells Augments Specific T-Cell Xenoresponse

The rejection of xenografts is characterized by infiltration of monocytes and natural killer (NK) cells into the graft, suggesting an important role for the innate immune system in xenorecognition. In this study, purified human NK or T cells were cocultured with porcine endothelial cells, and cytokines were analyzed by ELISA and intracellular FACS. We demonstrated a vigorous human anti-porcine xenoresponse that was associated with a strong T-cell proliferation against porcine endothelial cells. Limiting dilution cloning and T-cell receptor (TCR) Vbeta gene usage revealed a low number of xenoreactive T-cell precursors. We demonstrated that xenogeneic porcine but not allogeneic human endothelial cells induced the early production of interferon (IFN)-gamma by human NK cells but not by CD3+ T cells. Porcine xenoantigen-induced IFN-gamma production was only partially dependent on IL-12. Blocking IL-12 with neutralizing antibodies or by depletion of human macrophages partially decreased IFN-gamma production by CD56+ NK cells. Three-color flow cytometry revealed that IL-12 was produced through a species-specific activation of human macrophages by porcine endothelial cells. Our results indicate that the direct activation of NK cells and macrophages by porcine endothelial cells provides a unique pathway of xenorecognition that augments downstream specific T-cell immunity and represents a powerful effector mechanism in xenograft rejection.     

胚胎牌细胞不同移植途径对小鼠接种性肉瘤生长和自然杀伤细胞活性的影响

分别于肌肉、腹腔、周围静脉、门静脉对荷瘤小鼠移植胚胎牌细胞，并于移植后30天、60天分别测定瘤体积抑制率和自然杀伤细胞（NK细胞）活性。结果显示：上述4种移植途径在移植后30天均能明显抑制移植性肿瘤生长，NK细胞活性显著升高（P＜0．01）。但移植后60天，仅门静脉组对肿瘤生长保持良好的抑制状态，NK细胞活性保持较高水平，证明经门静脉移植是牌细胞移植最好的途径。     

Efficacy and Limitations of Natural Killer Cell Depletion in Cyclophosphamide-Induced Tolerance

Purpose We previously developed a cyclophosphamide (CP)-induced tolerance protocol, consisting of an intravenous injection of 1 × 10⁸ donor spleen cells (SC) given on day 0 and an intraperitoneal injection of 200 mg/kg CP given on day 2. In the present study, we modified this protocol with natural killer cell (NK) depletion in recipient mice, and evaluated the efficacy of tolerance induction. Methods We used B10.D2 (H-2^d; IE⁺) and B10 (H-2^b; IE⁻) mice as both donors and recipients. The recipient mice were treated with donor SC, CP, and donor bone marrow cells (BMCs) with or without NK depletion. Results A higher level of mixed chimerism was achieved in the NK-depleted recipients. Survival of both the skin and heart donor grafts was significantly prolonged in the NK-depleted recipients. Donor reactive Vβ11⁺ T cells were found at the same level as in untreated control mice. Pretreatment with recipient NK cell depletion was effective in inducing higher levels of donor mixed chimerism; however, permanent engraftment of donor bone marrow was not achieved. Conclusion Survival of donor grafts was remarkably prolonged in the NK cell-depleted group, but transplantation tolerance could not be induced. Our results suggest that NK cell depletion in CP-induced tolerance conditioning has some effect on the induction of donor-specific tolerance.     

Adoptive Immunotherapy of Patients with Metastatic Renal Cell Cancer Using Lymphokine-Activated Killer Cells, lnterleukin-2 and Cyclophosphamide: Long-Term Results

Background Initial results of adoptive immunotherapy using lymphokine-activated killer (LAK) cells and interleukin-2 (IL-2) appeared to offer promise for treating renal cell cancer (RCC). However, lower response rates were seen in subsequent trials, and the long-term results of this treatment method have not been fully reported. In this study, we examine long-term results of adoptive immunotherapy using LAK cells, IL-2, and cyclophosphamide (LAK/IL-2/CPM therapy). Methods We administered 10 courses of therapy to 9 patients with advanced RCC. One patient had liver and para-aortic lymph node metastases; the others had only lung metastases. The clinical effects were initially evaluated 4 weeks after therapy and follow-up was continued for periods of 43 to 76 months. Results The 4-week evaluation revealed 3 complete responses (CR), 3 partial responses (PR), 1 minor response (MR), 1 patient with no change in disease status (NC), and 2 patients whose disease progressed (PD). One CR patient remained apparently free of disease for 43 months. After tumors recurred in the lung of another CR patient further disease progression was suppressed by IL-2 administration until the patient died from other causes at 46 months. The third CR patient showed tumor recurrence in the lung and was re-treated with the sameLAK/CPM/IL-2 therapy. Lung tumors decreased in size (PR), but the patient died due to brain metastasis 2 months after the second round of treatment. The 2 initial PR patients, as well as the MR and NC patients, developed regrowth or new metastatic lesions within 2 to 15 months following therapy. The 2 PD patients died 2 and 9 months after therapy. Conclusion Long-term effects ofLAK/IL-2/CPM therapy were not correlated with the maximal response observed 4 weeks after therapy. AlthoughLAK/IL-2/CPM therapy appears suitable for use as induction therapy in RCC, our data suggest that long-term suppression will require surgical removal of remnant tumors or more intensive maintenance therapy.     

Antitumor Killer Lymphocytes in the Peripheral Blood of a Patient with Transitional Cell Carcinoma of the Bladder

Background: Peripheral blood lymphocytes (PBL) from patients with bladder cancer also contain cells possessing cytotoxic activity against autologous tumor cells. These cells are phenotypically heterogenous and include natural killer (NK) and cytotoxic T cells. This study investigated the role of cytotoxic lymphocytes directed against autologous bladder cancer cells.
Methods: PBL were obtained at intervals before and after surgery and analyzed for cytotoxic activity against autologous bladder cancer cells in 4-hour⁵¹ Cr release assay. PBL stimulated with autologous tumor cells were also transformed with human T-lymphotropic virus type-1, establishing a cell line (KB31) which was analyzed for phenotype and cytotoxic activity against the autologous tumor cells.
Results: PBL preoperative cytotoxic activity was low, but increased after surgery. Cytotoxic activity was found not only against autologous bladder cancer cells, but also against heterologous bladder cancer (KK-47) and myeloid leukemia (K562) cells, with the highest activity against the heterologous cell lines. The cytotoxic activity of KB31 was 40|X% against autologous tumor cells 6 weeks after initiation of the cell line, but decreased to 5|X% by 6 months. This activity was lower than that against the other cell lines, and was similar to that of PBL in short-term culture. Fluorescence-activated cell sorter (FACS) analysis demonstrated that in KB31 cells at 6 weeks, CD8+ cells were dominant, but CD56+ cells predominated at 6 months.
Conclusion: These results suggest that the presence of cytotoxic activity in the peripheral blood of the patient was due to both cytotoxic T cells and NK cells. The cytotoxic activity was lowest prior to surgery and increased postoperatively.     

Critical role of natural killer cells in the rejection of human hepatocytes after xenotransplantation into immunodeficient mice

The severe combined immunodeficiency/albumin linked‐urokinase type plasminogen activator (SCID/Alb‐uPA) human liver chimeric mouse model has added a new dimension to studies of liver based human diseases and has important potential for study of human hepatic drug metabolism. However, it remains unclear if natural killer (NK) cell in SCID/Alb‐uPA mice has an important negative impact on engraftment and expansion of human hepatocytes after transplantation. Here, we explore the role of mouse NK cells in the rejection of transplanted human hepatocytes in SCID/Alb‐uPA mice. We assessed NK cell activity in vivo, using ¹²⁵I‐iodo‐2′‐deoxyuridine incorporation assay. Low serum human alpha‐1 antitrypsin (hAAT, <10 μg/ml) recipients, representing graft failure, showed resistance to engraftment of MHC class I knockout marrow (indicating high NK cell activity), while NK cell‐depleted low hAAT recipients and high hAAT (>100 μg/ml) recipients accepted MHC class I knockout marrow, indicating a correlation between low NK cell activity, in vivo, and high level human hepatocyte engraftment. We also showed that higher level engraftment of human hepatocytes was achieved in both NK cell‐depleted SCID/Alb‐uPA mice and Rag2^?/?γc^?/?/Alb‐uPA (T,B and NK cell deficient) mice compared with untreated SCID/Alb‐uPA mice. These results support a critical role for mouse NK cells in the rejection of human hepatocytes xenotransplanted to immunodeficient mice.