Systemic administration of human leukocyte interferon to melanoma patients. III. Increased helper:suppressor cell ratios in melanoma patients during interferon treatment

Malignant melanoma patients treated with human leukocyte interferon (IFN-alpha) displayed increased natural killer (NK) cytotoxicity to K562 targets within the first 2 weeks of therapy. This study explored the possibility of T-cell regulation of this NK response, as evidenced by a variation in T-cell subpopulations. T-cell subset levels were studied in 9 patients who received daily doses of IFN-alpha over a period of 42 days. Five monoclonal antibodies to T-cell surface markers were used: Leu 1 (pan-T), Leu 2a (T suppressor/cytotoxic), Leu 3a (T helper/inducer), HNK-1 (Leu 7, a marker for NK cells), and B73.1 (an antibody against the Fc receptor on NK/K cells). Percentages of these markers were measured on days 0, 3, 7, and 21 of treatment. Percentages of Leu 1-positive cells and Fc-receptor-positive cells remained relatively constant throughout treatment in all patients. A trend toward a decrease in suppressor cells and an increase in helper cells peaking on day 7 and returning to earlier values by day 21 was found in 5 patients. The increase in NK cytotoxicity was not reflected consistently in proportions of HNK-1-positive cells or Fc receptor-bearing cells within the first week of therapy. The most significant finding was an increase in the helper:suppressor cell ratio peaking on day 7 and returning to pretreatment values by day 21. This increase was seen in every patient studied. The average pretreatment Leu 3a:Leu 2a ratio was 0.67 increasing to an average value of 1.47 on day 7 (p less than 0.005). Leu 3a:Leu 2a ratios returned to pretreatment values, in parallel to NK activity, despite continuation of interferon therapy.     

Recombinant interleukin-2 treatment in patients with metastatic colorectal cancer: Effect on natural cytotoxicity

Summary Natural cytotoxicity (natural killer, NK, and lymphokine-activated killer, LAK, activity) was documented in 12 patients with metastatic colorectal cancer, both before and after a 5-day course of continuous therapy with intravenous recombinant interleukin-2 (rIL-2). Treatment induced a substantial increase in circulating CD56⁺ lymphocytes (pretreatment: 12.1±6.9%, mean ± SD; posttreatment: 39.2±6.9%. Maximal NK cell activity was induced by treatment with rIL-2 but only suboptimal augmentation of LAK cell cytotoxicity was obtained. This study indicates that although continuous infusion of rIL-2 does have a significant effect on natural cytotoxicity, this is suboptimal and further studies are necessary to define the most efficacious immunity-enhancing regimens of therapy, thereby hopefully improving clinical outcome of rIL-2 treatment.     

Modulation of natural killer cytotoxicity by muramyl dipeptide and trehalose dimycolate incorporated in squalane droplets

Summary The effect on natural killer (NK) cytotoxicity of splenic cells from BALB/c mice pretreated i. v. with squalane-in-water preparations of muramyl dipeptide (MDP), trehalose dimycolate (TDM), or the combination of MDP-plus-TDM was investigated. MDP or TDM augmented the NK cytotoxicity which peaked 48 h after the pretreatment whereas the combination of MDP and TDM induced an inhibition of the NK activity. Infection with influenza virus, a potent stimulator of NK cells, after the pretreatment with biological response modifiers resulted in a markedly enhanced NK activity on day 2 in MDP and control groups. Mice pretreated with TDM or the combination of MDP and TDM showed only moderate NK activity which peaked on day 3 after influenza infection. The NK activity was susceptible to asialo GM₁ and complement treatment. The cytotoxicity of MDP-plus-TDM cells could be significantly enhanced after treatment with anti-macrophage monoclonal antibody and complement. NK activity induced by MDP or TDM was reduced by mixing MDP-plus-TDM cells. Addition of adherent cell-depleted MDP-plus-TDM suspension to MDP or TDM cells had a NK restorative effect. Splenic cells from mice pretreated 2 days earlier with MDP or TDM, but not MDP-plus-TDM, generated enhanced levels of luminol-dependent chemiluminescence.     

Selenium and Glutathione Peroxidase Status in Adult Egyptian Patients with Acute Myeloid Leukemia

This study was undertaken to evaluate selenium (Se) and glutathione peroxidase (GPX) status in patients with newly diagnosed acute myeloid leukemia (AML) before and after induction therapy. Twenty-five patients with newly diagnosed AML and 15 healthy age- and sex-matched control subjects were included in this study. Serum Se level by the graphite furnace atomic absorption spectrometric technique and GPX activity by an adaptation of Beutler method was performed for the patients before and after receiving the induction therapy. Serum Se level was significantly lower in patients with AML versus control subjects (63.1?±?8.8 versus 77?±?8.8 µg/L before therapy with a P value <0.01 and 69?±?6.8 versus 77?±?8.8 µg/L after therapy with a P value <0.01).GPX activity was significantly lower in patients with AML versus control subjects (1.6?±?0.4 versus 3.4?±?0.7 µ/g protein pretreatment with a P value <0.01and 1.9?±?0.6 versus 3.4?±?0.7 µ/g protein post induction treatment with P value <0.01).Se level and GPX activity significantly increased in AML patients after treatment. Patients who accomplished complete remission after induction harbored significantly higher Se levels than resistant patients before and after treatment. There was no significant correlation between serum Se level and GPX activity. Decreased Se level and reduced GPX activity in AML patients support the association of carcinogenesis and subnormal Se states.     

Activity of natural cytotoxic cells in non-Hodgkin's lymphoma]

Cytotoxic activity of NK cells in the peripheral blood has been determined in 30 patients with malignant non-Hodgkin lymphomas prior to and following therapy. In the whole group as well as in subgroups classified according to the criteria of Working Formulation as lymphomas of the low, moderate and high degree of malignancy, activity of NK cells has been statistically significantly lower than that in healthy individuals. Marked increase in this activity has been noted in 19 patients in the state of clinical remission after the treatment with cytotoxic agents, and sometimes radiotherapy. The value of mean cytotoxic activity reached normal limits in the lymphoma of high degree of malignancy, and exceeded these limits in the lymphomas of moderate and low malignancy.     

Specific immunotherapy with vaccinia oncolysates

Summary Short- and long-term effects of IV administration of human fibroblast interferon (HFIF) on natural cytotoxicity was studied in patients with HBsAg-positive chronic active hepatitis. Short-term kinetics demonstrated a transient decrease of natural cytotoxicity, when measured 2 or 4 h after IV administration of HFIF (1–10×10⁶ U/injection). Twenty-four hours after the initial injection of HFIF natural cytotoxicity was increased to 196%–282% of pretreatment values. The kinetics of NK activity during chronic stimulation with HFIF revealed the following features: (a) The highest relative increase was seen during the initial phase of HFIF application; (b) enhanced NK activity could be maintained for 2–4 weeks of therapy; (c) at a plateau of high activity short-term increases were much less pronounced; (d) in all patients monitored so far over a period of several weeks a gradual decrease of augmented NK activity has been observed despite continued administration of high doses of HFIF.These findings indicate that in vivo administration of HFIF results in an augmentation of NK activity in man. Prolonged treatment with HFIF seems to exhaust the NK cell system. Monitoring of natural cytotoxicity may be of critical importance for the determination of an administration schedule of interferon for future therapy.     

Human epidermal cells and squamous carcinoma cells synthesize a cytokine that augments natural killer cell activity

Normal as well as transformed epidermal cells (EC) have recently been reported to produce a cytokine--EC-derived thymocyte-activating factor (ETAF), which according to its biologic as well as biochemical properties is indistinguishable from macrophage-derived interleukin 1 (IL 1). In the present study, the effect of supernatants (SN) derived from normal EC and a human squamous carcinoma cell (SCC) line were tested for their effects on natural killer (NK) cell activity. EC- as well as SCC-derived SN were able to augment in vitro NK cell activity of peripheral blood lymphocytes against K 562 cells. In contrast, adherent cell-derived, IL 1-containing SN did not affect NK cell activity. Upon high-pressure liquid chromatography (HPLC) gel filtration, ETAF and the EC-derived NK cell activity-augmenting factor (ENKAF) exhibited a similar m.w. However, by using reverse-phase HPLC, ETAF and ENKAF eluted as distinct peaks of activity, indicating that SCC cell-derived ENKAF is different from ETAF. Furthermore, ENKAF does not contain interleukin 2 (IL 2) or interferon (IFN) activity. The enhancement of NK cell activity was dose dependent and evident after 20 hr of preincubation of effector cells. Pretreatment of target cells with ENKAF did not affect the susceptibility of the target cells. The NK activity of large granular lymphocytes (LGL) purified by discontinuous Percoll gradient centrifugation and further depleted of high-affinity sheep erythrocyte rosetting cells was enhanced by ENKAF. In contrast, no NK cell activity was expressed by LGL-depleted T cell populations before or after treatment with ENKAF. In a single cell cytotoxicity assay in agarose, the number of lymphocyte binding to K 562 was not affected by ENKAF, but the frequency of dead conjugated target cells and presumably of active killer cells was increased by pretreatment with ENKAF. Additional incubation of LGL with ETAF did not further increase ENKAF-mediated augmentation of NK activity. In contrast to ETAF, ENKAF was not chemotactic for polymorphonuclear leukocytes. These results indicate that normal as well as transformed EC release a unique cytokine--ENKAF--which augments NK cell activity of LGL but is distinct from ETAF, IL 2, and IFN.     

Effect of chemotherapy on NK function in the peripheral blood of cancer patients

Summary The effects of cytotoxic chemotherapy on NK cell function and on glass adherent cell regulation of NK cell function was evaluated in the peripheral blood mononuclear cells of 20 previously untreated solid tumor patients. Most of the patients studied had lung cancer and received one of four combination chemotherapy treatment regimens. In addition, one patient with colon carcinoma and one patient with melanoma were studied, each of whom received treatment with a single agent. The results demonstrated that chemotherapy exerted a differential influence on NK activity which correlated with the pretreatment NK level of function in the individual patient. In patients with depressed NK levels prior to treatment, chemotherapy augmented NK function; in patients with normal levels prior to treatment, chemotherapy depressed NK function. The effects observed appeared to be associated with the capacity of chemotherapy to influence glass adherent cell regulation of NK activity. There was no apparent correlation between the effects of chemotherapy on numbers of NK effector cells, Leu11+ cells, or latex-ingesting cells. Also, there was no correlation between the effects seen and the type of drug treatment that was administered; rather, this was dependent on the pretreatment NK level of function which in turn was associated with glass adherent cell regulation of NK function.Supported in part by PHS Grant No. 27598     

感染性心内膜炎与肠道菌群的相关性研究

目的 探讨感染性心内膜炎与肠道乳杆菌、双歧杆菌和大肠埃希菌的临床相关性。方法 选择2008年4月至2016年4月湖州市第一人民医院心内科病房收治的单纯感染性心内膜炎患者67例为观察组,选择同期来院体检结果正常者65例为对照组。分别收集观察组患者治疗前1天、治疗第3天和治疗1周后3个不同时间节点的粪便标本,同时收集对照组的粪便标本,通过FISH实验检测粪便中双歧杆菌和乳杆菌的含量差异,同时通过16SrRNA/DNA荧光定量PCR技术进一步明确观察组患者3个时期肠道双歧杆菌、大肠埃希菌和乳杆菌与对照组的数量差异。结果 观察组患者治疗前1天双歧杆菌值为6.2%±0.56%,治疗第3天为4.3%±0.34%,治疗1周后为8.7%±0.56%,对照组双歧杆菌值为9.4%±0.98%。观察组患者治疗前1天乳杆菌值为5.4%±0.86%,治疗第3天为4.9%±0.24%,治疗1周后为6.1%±0.72%,对照组乳杆菌值为8.3%±0.51%。治疗前1天与治疗1周后,观察组患者双歧杆菌和乳杆菌值与对照组相比,差异均有统计学意义（P<0.05）。16SrRNA/DNA荧光定量PCR检测表明,观察组患者在3个治疗时期中肠道乳杆菌、双歧杆菌和大肠埃希菌数量的对数值与对照组比较差异均有统计学意义（P<0.05）。其中观察组患者双歧杆菌和乳杆菌数量在治疗第3天较低,1周后有所回升;大肠埃希菌数量在治疗第3天亦较低, 但1周后没有明显回升。结论 感染性心内膜炎患者肠道菌群数量与健康人相比有所下降,肠道乳杆菌、大肠埃希菌和双歧杆菌失调可能影响其发病和转归。     

The distribution of the major peripheral blood T, B and NK cell subsets does not predict the clinical outcome of Graves' disease patients after methimazole therapy

Biological markers capable of predicting the clinical outcome of antithyroid drug therapy could be clinically useful in selecting the modality of treatment for Graves' disease, but at present they are unavailable. In the present study we prospectively explore the value of 22 different peripheral blood T, B and NK lymphocyte subsets to predict remission and relapse in a group of 42 Graves' disease patients. Eighteen patients were studied at diagnosis, before treatment, and 24 during antithyroid drug therapy. All cases were followed-up for at least one year after finishing an 18 month cycle of methimazole therapy. The combination of flow cytometry and 3- color immunofluorescence did not reveal significant differences in the distribution of the major peripheral blood T, B and NK cell subsets between the relapsed patients and those in remission, both in the groups studied at diagnosis and in those analyzed during the cycle of antithyroid drug therapy. In our search for a prognostic marker for relapse prediction we found that some lymphoid subpopulations such as total B cells, total NK and NK CD8+ cells showed high sensitivity (88-100%). In turn, other subsets such as TCD8+, total T and B cells expressing the CD25 antigen displayed high specificity (77-88%).     

Effects of metabolic inhibitors on spontaneous and interferon-boosted human natural killer cell activity

Human natural killer (NK) cell activity can be augmented by pretreatment with partially purified preparations of human interferon (IF). Studies have now been performed to determine the metabolic processes required for and involved in spontaneous NK activity and augmentation of cytotoxicity. A 4-hr 51Cr release cellular cytotoxicity assay was used to measure the NK activity, and peripheral blood leukocyte cells (PBL) were treated with: a) x-ray or mitomycin C; b) actinomycin D; or c) emetine, cycloheximide, pactamyhcin, or puromycin to assess the roles of DNA, RNA, and protein synthesis, respectively, in spontaneous NK activity and in boosting by IF. Prolonged incubation (18 hr) of PBL after blockage of synthesis of DNA almost completely abrogated NK activity; however, NK activity could be partially or totally restored to these populations by incubation of the effector cells for 1 hr at 37 degrees C with IF. Blockage of DNA synthesis for 1 hr had no effect on spontaneous NK activity or on boosting by IF. Inhibition of RNA synthesis also had no effect on spontaneous NK activity. Treatment of PBL with actinomycin before exposure to IF prevented boosting, but treatment with the RNA synthesis inhibitor after boosting with IF for 5 to 6 hr no longer had an appreciable effect on cytotoxicity. The effect of protein synthesis inhibitors on spontaneous NK activity was dependent on the inhibitor selected. Emetine and puromycin totally abrogated spontaneous NK activity at concentrations of inhibitor that blocked 3H-leucine incorporation 90% or more. In contrast, cycloheximide and pactamycin had only minimal effects on spontaneous NK activity but totally abrogated the boosting of IF.     

Decreased natural killer activity in thalassemia major: a possible consequence of iron overload

We have investigated the natural killer (NK) activity of both fractionated (Percoll density gradient) and unfractionated mononuclear cells from patients with beta-thalassemia major who are iron overloaded as a consequence of chronic transfusion therapy. These patients were found to have significantly decreased NK activity against K562 targets at all effector:target ratios tested (p less than 0.001). Both splenectomized and nonsplenectomized patients had normal proportions of Leu-11b-staining (NK) cells. Since they had normal to elevated absolute white cell and lymphocyte counts, a change in the absolute number of NK cells could not account for the decreased killing. We also found that the decrease in NK activity was transfusion related (r = -0.603, p less than 0.005). To determine whether or not this decrease in NK activity could be related to iron overload, we preincubated patient effector cells with desferrioxamine (DFO) or 2,3-dihydroxybenzoic acid (DHB) for 6 hr before addition of K562 targets. Both of these iron-chelating agents consistently increased the NK activity of cells from thalassemia patients. DHB had the greater effect, being able to increase patient NK activity to virtually normal levels. On the other hand, preincubation of cells from normal controls with DHB caused only a slight increase in NK activity, while similar treatment with DFO had little or no effect. When target cells were preincubated with the chelating agents before addition of either normal or patient effector cells, no change in cytotoxicity was seen, demonstrating that the chelating agents act at the effector cell level. Furthermore, if the chelating agents were saturated with iron prior to preincubation with the effectors, no increase in the cytotoxicity of thalassemic NK cells was observed. These results indicate that thalassemia patients have a reversible, transfusion-related decrease in NK function which may arise as a consequence of iron overload.     

Multicenter clinical trial to evaluate the therapeutic use of recombinant growth hormone from mammalian cells in the treatment of growth hormone neurosecretory dysfunction

The efficacy and safety of a 12-month treatment with recombinant human growth hormone from mammalian cells (r-hGH, Saizen) in growth hormone neurosecretory dysfunction (GHND) are evaluated in this study. r-hGH was administered subcutaneously, at a dosage of 0.5 IU/kg/week divided into 6 equal daily doses. A total of 16 (12 M and 4 F) poorly growing patients, height -2.3 SD or more below the mean for chronological age and sex, were included in the study. r-hGH therapy significantly increased the growth velocity; from 3.57 +/- 0.85 cm/year, before therapy, to 7.09 +/- 2.29 cm/year after 12 months (p less than 0.001). Patients' height SD score rose from -3.40 +/- 0.84 SDS to -2.98 +/- 0.69 SDS (p less than 0.01). Somatomedin C increased significantly from a baseline value of 0.59 +/- 0.32 U/ml to 1.26 +/- 0.66 U/ml after therapy (p less than 0.01). Finally, r-hGH therapy improved the pretreatment adult height prediction; from an initial prognosis of -2.66 +/- 0.79 SDS to -2.17 +/- 0.81 SDS after treatment (p less than 0.01). No side effects or adverse reactions were observed during treatment. Anti-r-hGH antibody formation was not found in any of the patients included in the study.     

Decrease in the phagocytic activity of peripheral monocytes in patients treated with human interferon-α

Summary The influence of daily intramuscular injections of 3×10⁶ units of interferon- (IFN-) on the phagocytic activity of peripheral monocytes was studied in 28 tumor patients. One day after initiation of IFN therapy no major change in the capacity of monocytes to ingest yeast particles was observed. After 1 week, 3 months, 6 months, and 9 months of treatment, monocyte phagocytosis had decreased in the majority of the patients tested. In patients where natural killer (NK) cell activity was measured simultaneously with monocyte phagocytosis, a correlation between the degree of enhancement of NK activity and the degree of decrease in monocyte phagocytosis was observed.     

腹腔镜全结肠系膜切除术治疗老年结肠癌患者的近期疗效评价及对机体免疫力的影响

目的:研究腹腔镜全结肠系膜切除术对老年结肠癌患者的近期疗效评价及机体免疫力的影响,为临床治疗结肠癌提供依据。方法:选择2014年6月至2016年6月我院收治的136例老年结肠癌患者为研究对象,按照患者治疗意愿分为两组。治疗组57例行腹腔镜下全结肠系膜切除术,对照组79例行开腹手术。比较两组患者的手术时间、淋巴结清扫数量、术中出血量、肛门排气时间、下床活动时间、术后住院时间及术后并发症发生率。于术前1 d、术后1 d及术后3 d采用流式细胞仪检测两组患者外周血中T淋巴细胞亚群(CD3~+、CD4~+、CD8~+)及自然杀伤(NK)细胞;于术前1 d、术后1 d及术后3 d采用酶联免疫吸附法(ELISA)测定两组患者血清C反应蛋白(CRP)、可溶性白细胞介素-2受体(sIL-2R)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)浓度。结果:与对照组比较,治疗组术中出血量减少,肛门排气时间、下床活动时间及术后住院时间均缩短,差异均有统计学意义(P0.05)。两组患者淋巴结清扫数量及手术时间比较,差异无统计学意义(P0.05)。治疗组并发症发生率为7.02%,低于对照组的17.72%,差异具有统计学意义(P0.05)。治疗组患者术后1 d和术后3 d外周血中T细胞亚群CD3~+、CD4~+和NK细胞所占比例及CD4~+/CD8~+的比值与术前1 d相比有所减小(P0.05),且术后1 d的CD4~+/CD8~+的比值和NK细胞低于术后3 d(P0.05);两组术后1 d及术后3 d外周血中CD8~+所占比例与术前1 d有所升高,但是差异无统计学意义(P0.05),且术后1 d的CD8~+所占比例与术后3d比较无明显差异(P0.05)。治疗组患者术后1 d和术后3天外周血中CD3~+、CD4~+与对照组比较,差异无统计学意义(P0.05),CD4~+/CD8~+的比值和NK细胞高于对照组,差异有统计学意义(P0.05)。治疗组术后1 d及术后3 d血清CRP、sIL-2R、IL-6、IL-8水平与均较术前1 d增高,术后3 d治疗组血清中CRP、sIL-2R、IL-6、IL-8水平低于术后1 d(P0.05),治疗组术后1 d及术后3 d血清中CRP、sIL-2R、IL-6、IL-8水平均低于对照组,差异均具有统计学意义(P0.05)。结论:腹腔镜全结肠系膜切除术与开腹手术相比,近期疗效更好,具有创口小、出血量少、术后恢复快、术后并发症发生率低及对机体免疫功能影响小等优点,可应用于老年结肠癌的治疗。     

Interleukin 2 enhances the natural killer cell activity of acquired immunodeficiency syndrome patients through a gamma-interferon-independent mechanism

Patients with the acquired immunodeficiency syndrome (AIDS) exhibit a variety of disorders of cellular immunity, including a deficient ability to generate cytotoxic T cells and depressed levels of natural killer (NK) cell activity. Interleukin 2 (IL 2) in vitro can markedly augment these depressed immune functions. Because IL 2 can induce the release of interferon-gamma (IFN-gamma) from normal peripheral blood lymphocytes (PBL), and because IFN-gamma may play a role in the regulation of NK cell activity, this study was performed to determine if the IL 2 enhancement of the NK cell activity of patients with AIDS was an IFN-gamma-dependent effect. PBL from eight healthy heterosexual donors and from nine patients with AIDS were studied for their ability to release IFN-gamma in response to IL 2 at a concentration of 100 U/ml. After 60 hr of culture, the PBL of all eight healthy donors produced IFN-gamma with a mean titer of 113 U/ml (range 40 to 320 U/ml). In contrast, the PBL from only two of nine patients with AIDS released measurable amounts of IFN-gamma (40 U/ml each) in response to IL 2 with a mean titer of 13.5 U/ml for all nine. Although the PBL from patients with AIDS were deficient in their capacity to produce IFN-gamma in response to 100 U/ml of IL 2, significant enhancement of NK cell activity could be obtained after only 1 hr of PBL treatment with 10 U/ml of IL 2, with an optimal NK enhancing effect occurring at doses of 50 to 100 U/ml of IL 2. The use of an anti-IFN-gamma monoclonal antibody resulted in complete neutralization of the IFN released from the normal PBL cultured with IL 2, but failed to inhibit the IL 2 enhancement of NK cell activity. Exogenous IFN-gamma exhibited different kinetics of enhancement of NK cell activity when compared to IL 2, requiring substantially more than 1 hr of pretreatment of PBL. These results indicate that the PBL from patients with AIDS usually do not release IFN-gamma when cultured with IL 2, and that IL 2 enhancement of the depressed NK cell activity of these patients may be an IFN-gamma-independent event. These results may have important implications for the therapy of AIDS.     

COUMARIN THERAPY—Prothrombin Activity after Termination of Treatment

Twelve patients receiving coumarin type hypoprothrombinemic agents were studied before, during and after termination of therapy, the prothrombin proconvertin method having been used to assay the prothrombin activity complex.In no instance was post treatment “rebound” demonstrated.Prothrombin activity levels returned to pretreatment values only after ten days following termination of coumarin or Dicumarol administration.If a reactivation of thrombotic tendency occurs following discontinuance of anticoagulant therapy, it would not appear to be related to a “rebound” of prothrombin activity above that which is “normal” for the individual patient.Patients tend to return to the same level of prothrombin activity present before initiation of coumarin therapy.     

G-CSF-induced evacuation of sinusoidal NK cells and the facilitation of liver regeneration in a partial hepatectomy

Since liver regeneration after partial hepatectomy (PHx) is known to improve by pretreatment with recombinant human G-CSF (rhG-CSF), we investigated the mechanism by evaluating the distribution and activity of sinusoidal NK cells. F344 rats were treated with rhG-CSF (250 microg/kg/day) for 5 days before PHx. Pretreatment with rhG-CSF improved the serum ALT levels and DNA biosynthesis of the remnant liver tissues at 20 h after PHx. Notably, the rhG-CSF pretreatment decreased the number of NK cells in the liver determined by immunohistochemistry using anti-NKR-P1A mAb before and at 20 h after PHx with no significant change in the NK activity per cell base, while also increasing the number of NK cells in the peripheral blood detected by flow cytometry. The rhG-CSF induced a pre-PHx downregulation of the IL-12p70 protein levels, while also promoting the post-PHx reduction of the protein levels of IL-12p70 and IFN-gamma. Conversely, rhG-CSF had no effect on the pre-PHx mRNA levels or the PHx-induced upregulation of mRNA levels of TNF-alpha, IL-1beta, IL-6, TGF-beta, IL-10, HGF, and c-Met determined by real-time RT-PCR. These results strongly suggest that rhG-CSF-induced facilitation of liver regeneration is achieved by immunoregulation through the intrahepatic IL-12 downregulation and evacuation of sinusoidal NK cells.     

Evidence for a beta-adrenoceptor-mediated regulation of human natural killer cells

Pretreatment of lymphocytes (16 hr, 37 degrees C) with adrenaline at final concentrations of 10(-7) to 10(-9) M, followed by removal of the drug, increased natural killer (NK) cell activity vs K562 leukemic cells in a 4-hr 51Cr-release assay. The most efficient concentration of adrenaline was 10(-8) M; mean increase of NK activity over base-line activity for all donors examined was 30%. However, the individual response to adrenaline pretreatment was variable; in some donors, the effect was equal to maximal interferon (IFN) stimulation. Effects of adrenaline pretreatment were consistently reduced to base-line activity by co-incubation with the nonselective beta-adrenoceptor antagonist propranolol at 100-fold higher concentrations. The enhancing effect of adrenaline (10(-8) M) pretreatment was also observed after 1-hr pretreatment; this effect was prevented by simultaneous incubation with propranolol but was not affected by dex-propranolol. Direct addition of adrenaline to lymphocyte/target cell mixtures was inhibitory at 10(-6) M adrenaline concentration. The inhibitory effect of adrenaline in this assay was again completely prevented by propranolol and unaffected by dex-propranolol. The observed stimulatory effect of adrenaline pretreatment could not be ascribed to IFN production. Data presented indicate a dual effect of adrenaline on NK cell activity and suggest both a positive and a negative beta-adrenoceptor-mediated regulation of human NK cells.     

Histological change after interferon therapy in chronic hepatitis C in view of iron deposition in the liver

We examined the efficacy of interferon (IFN) therapy for chronic hepatitis C (CHC) in view of the change of liver histology and iron staining before and after IFN therapy. Enrolled in this study were 109 patients with CHC who completed IFN treatment and were followed for at least 1 yr after the end of IFN therapy. Serum iron, unsaturated-iron-binding capacity (UIBC), and total-iron-binding capacity (TIBC) were assessed before IFN therapy. Knodell’s histological activity index (HAI) score and iron staining were examined in 55 patients in whom liver biopsy was performed at two points: before and 1 yr after IFN therapy. Serum iron levels before IFN therapy did not correlate with the response to IFN. The HAI score significantly decreased after IFN therapy in complete responders (p<0.01) and biochemical responders (p<0.01). Three factors in the HAI, periportal necrosis, intralobular necrosis, and portal inflammation, but not fibrosis, were significantly decreased in complete responders (p<0.01) and biochemical responders (p<0.01). Of 55 patients, 23 (41.8%) were positive for iron staining before IFN therapy and 14 of 55 (25.5%) after IFN therapy. The positive rate for iron staining tended to decrease after IFN therapy, not correlating to the response to IFN, but the change was not statistically significant. In conclusion, the histological improvement by IFN therapy was mostly seen in necroinflammatory changes but not in fibrosis at least 1 yr after IFN, and iron staining tended to decrease after IFN therapy.