M-CSF、IL-10对单核细胞IL-12、IL-18产生及HLA-DR和CD80表达的影响

目的 :探讨巨噬细胞集落刺激因子 (M CSF)和白细胞介素 10(IL 10 )对人外周血单核细胞分泌IL 12、IL 18及细胞表面HLA DR和CD80表达的影响。方法 :从健康献血员血液中分离单核细胞并进行体外培养 ,分别以M CSF和IL 10单独及共同作用 ,收集上清 ,用ELISA法检测单核细胞IL 12和IL 18的分泌 ,用流式细胞术检测细胞表面HLA DR及CD80的表达。结果 :①M CSF能诱导单核细胞分泌IL 18(P <0 .0 5 ) ;IL 10则能抑制IL 18的产生 (P <0 .0 5 ) ,并拮抗M CSF增强LPS诱生IL 18的作用 (P <0 .0 5 )。M CSF和IL 10均能抑制单核细胞分泌IL 12 p4 0 (P <0 .0 5 ) ,并具有协同效应 (P <0 .0 5 )。②M CSF能诱导单核细胞表面HLA DR的表达 (P <0 .0 5 ) ;IL 10则可抑制HLA DR的表达 ,并拮抗M CSF对HLA DR的诱导作用。M CSF对CD80表达的影响不大 ,IL 10能促进CD80的表达。结论 :M CSF和IL - 10可通过对单核细胞分泌IL 12、IL 18及HLA DR和CD80表达的调节 ,影响T细胞的活化、分化及其介导的免疫应答     

IL-6协同M-CSF体外诱导CD14~+单核细胞向M2样巨噬细胞分化

目的探讨IL-6协同M-CSF体外诱导CD14~+单核细胞向M2样巨噬细胞分化的作用机制,并检测诱导而来的M2样巨噬细胞的表型特点以及对肿瘤细胞的作用。方法体外构建IL-6诱导单核细胞分化模型,q RT-PCR检测M2表型分子(IL-10、TGF-β)在不同质量浓度IL-6刺激下的表达水平;Western blot检测STAT3信号通路的活化水平;采用细胞迁移、增殖实验验证IL-6诱导的M2样巨噬细胞对肿瘤细胞增殖、迁移的作用。结果在IL-6诱导分化的巨噬细胞中,IL-10、TGF-β表达增高呈IL-6剂量依赖性,且IL-6大量激活JAK/STAT3信号通路中的STAT3蛋白,使其磷酸化,si RNA沉默STAT3后,IL-10、TGF-β表达水平相应降低;此外,M2样巨噬细胞上清处理的实验组比对照组胃癌细胞增殖速度和迁移能力显著增强。结论 IL-6可以通过激活STAT3信号通路诱导单核细胞高表达IL-10、TGF-β,低表达IL-12的M2样巨噬细胞分化,并且M2样巨噬细胞可以促进肿瘤细胞的增殖和迁移。     

结核特异性多肽E6、E7和C14对单核-巨噬细胞亚型极化的影响

目的运用流式细胞术检测单核-巨噬细胞极化亚型M1(CD14~+CD16~+)及M2(CD14~+CD163~+),探讨结核特异性多肽对单核-巨噬细胞极化分型的影响。方法以3种结核特异性多肽E6、E7和C14作为刺激物,刺激人急性白血病单核细胞株(THP-1细胞)、结核病患者胸水单个核细胞及外周血单个核细胞(PBMC),用流式细胞术检测刺激物不同时间点对单核-巨噬细胞极化表面标记表达的影响。结果结核特异性多肽E6刺激THP-1 24+0 h、24+24 h、24+48 h、24+72 h后,CD16/CD163阳性率分别为16.85%/13.78%、19.59%/15.68%、18.14%/14.19%、13.61%/11.47%。E7刺激效果与E6相似,C14对THP-1的CD16/CD163表达影响不如前两种多肽强。结核特异性多肽E6刺激结核患者胸水或血标本单核细胞19 h后,CD14+CD16+阳性率(1#患者:1.66%,2#患者:4.37%)与CD14~+CD163~+阳性率(1#患者:1.76%,2#患者:2.82%)差异不大,但CD14~+CD16~+CD86~+阳性率(1#患者:2.20%,2#患者:6.16%)大于CD14~+CD163~+CD206~+阳性率(1#患者:1.37%,2#患者:0.92%)。E7刺激效果与E6相似,C14对结核患者胸水或血标本单核细胞CD14/CD16/CD163表达影响不如前两种多肽强。结论 3种结核特异性多肽特别是E6、E7体外刺激单核细胞株THP-1、结核患者胸水或血标本单核细胞主要向M1型单核-巨噬细胞极化。     

SRPX2经uPAR调控 人单核细胞系THP-1来源巨噬细胞迁移及M2极化

目的评价SRPX2蛋白对人单核细胞系THP-1来源巨噬细胞迁移及极化功能的影响。方法经佛波酯(PMA)诱导人单核细胞系THP-1为巨噬细胞后,将SRPX2重组蛋白作用于巨噬细胞,Transwell法检测细胞迁移,免疫荧光法检测SRPX2与uPAR的定位,Western blot检测相应信号通路蛋白的表达。再用IFN-γ及LPS诱导巨噬细胞的M1极化,SRPX2重组蛋白作用后,反转录PCR检测M1/M2标志物表达。结果 SRPX2明显促进人单核细胞系THP-1来源巨噬细胞的迁移(P0.01),加入uPAR中和抗体后,明显抑制迁移(P0.01)。在诱导M1极化的巨噬细胞中,经SRPX2重组蛋白作用后,M1标志物CD40和IL-6明显下降,而M2标志物CD206和IL-6明显上升(P0.01)。SRPX2与uPAR及CD11b的表达存在共定位。SRPX2重组蛋白作用后巨噬细胞FAK及Akt磷酸化水平增高。结论 SRPX2可能通过uPAR/CD11b/FAK/Akt通路促进人单核细胞THP-1来源巨噬细胞的迁移与M2极化。     

4-1BBL逆向信号促进单核细胞的增殖和分泌细胞因子的作用

采用抗人 4 1BBL单克隆抗体 1F1包被 2 4孔培养板 ,加入经免疫磁珠阴性选择获得的高纯度人外周血单核细胞 ,动态观察细胞的生长状态 ,并用3 H TdR掺入法分析单核细胞增殖 ,应用ELISA动态测定培养上清中IL 6、IL 10、M CSF和FL等细胞因子水平。结果发现 1F1非常有效地促进单核细胞的增殖 ,1F1组单核细胞分泌高水平的M CSF以及IL 6和FL增加 ,但 1F1组单核细胞分泌IL 10水平低于对照组 (P <0 0 5 )。由此表明 ,抗人 4 1BBL单克隆抗体 1F1通过激发单核细胞 4 BBL逆向信号 ,介导单核细胞分泌M CSF、IL 6和FL。上述细胞因子联合作用 ,从而促进了单核细胞的增殖。     

氧化修饰低密度脂蛋白对人外周血单核细胞和THP-1细胞功能的影响

以天然低密度脂蛋白(nLDL) 为对照组, 探讨了氧化修饰低密度脂蛋白(oxLDL)对人外周血单核细胞(PBMs) 和人前单核白血病细胞株THP 1 粘附和分泌功能的影响。结果表明: oxLDL可上调单核细胞粘附分子CD11b 表达, 促进其与内皮的粘附, 并进一步诱导TNF α、IL 8 的分泌; 而nLDL无此作用。说明oxLDL是单核细胞粘附和激活的诱导剂, 经oxLDL诱导的THP 1 表现出与成熟单核 巨噬细胞相似的特征, 可作为体外动脉粥样硬化单核细胞分化激活的细胞模型     

HBsAg体外冲击的慢性乙肝患者树突状细胞的生物学特性及其对HBV特异性CTL的诱导作用

目的 :研究HBsAg冲击的慢性乙肝患者单核细胞来源的树突状细胞 (DCs)的功能状况及体外对HBV特异性CTL的诱导作用 ,初步探讨诱导特异性抗HBV细胞免疫的途径。方法 :分离慢性乙肝患者外周血单核细胞 ,以GM CSF +IL 4 +TNF α培养诱导DCs,加入HBsAg冲击以诱导HBV特异性DCs。采用FCM测定细胞表面免疫分子CD1a、CD83、CD86、CD80、CD4 0以及HLA DR的表达水平 ,ELISA法检测培养上清中细胞因子IL 6、IL 12的分泌含量 ,MTT法测定DC刺激同种异体淋巴细胞增殖的能力 ,LDH法检测DC诱导的患者外周血T细胞对HepG2 2 2 15 (转染HBVDNA)、HepG2肝癌细胞株及K5 6 2白血病细胞株的细胞毒作用。结果 :HBsAg冲击的DC其表达CD1a、CD83、CD86、CD80、CD4 0、HLA DR表面分子明显高于对照组 (P <0 0 1,P <0 0 5 ) ,分泌IL 12的水平也高于对照组 (P <0 0 1) ,而分泌IL 6的水平则较对照组显著降低(P <0 0 1) ;HBsAg冲击的DC刺激同种异体淋巴细胞增殖的能力明显增强 (P <0 0 5 ) ,并可有效地诱导自体CTL对转HBV基因的HepG2 2 2 15细胞高效特异性杀伤作用 (P <0 0 1)。结论 :慢性乙型肝炎患者单核细胞来源的DCs经HBsAg抗原冲击后 ,生物学活性增强 ,并且能有效地诱导对HBV特异性反应的CTL。     

多参数流式细胞术鉴别诊断单核细胞相关性白血病的意义

目的：用多参数流式细胞术（FCM）鉴别诊断单核细胞相关性白血病（MLIL）。方法：采用CD45／SSC散点图设门多参数流式细胞术分析179例白血病细胞CD14及其它相关抗原的表达情况。结果：CD14在179例白血病中的表达，AML25．6％（30／117），CML5．9％（1／17）。CMML100％（3／3）；ALL、CLL、AUL、BAL CD14都阴性。117例AML中，AML-M4 CD14阳性占75．0％（15／20），AML-M5a 22．2％（2／9），AML-M5b 92．9％（13／14）。CD14在AML-M4／M5的阳性率无明显差异（P=0．486），而AML-M5a／M5b中有显著性差异（P〈0．001）。AML-M0、M1、M2、M6、M7中CD14全部阴性。在AML组中CD14的表达与CD117、CD34呈负相关。与CD64、HLA-DR、CD4、CD36、CD15、CD11b呈正相关。在CD45／SSC、CD71／CD33、CD15／CD11b散点图中M4／M5的粒细胞和单核细胞克隆位点明显不同。结论：CD14对单核细胞相关性白血病具有高特异性，敏感性不足；联合其它抗体不但可以区别单核细胞相关性白血病，而且可以鉴别AML-M4／M5亚型；CD45／SSC、CD71／CD33、CD15／CD11b三幅散点图有助于鉴别粒细胞和单核细胞克隆。     

炙甘草多糖对小鼠巨噬细胞再极化的影响

目的:建立探讨炙甘草中等分子量多糖(MPRR)诱导巨噬细胞极化的情况,阐明肿瘤微环境中极化后巨噬细胞对小鼠4T1乳腺癌细胞生长的影响。方法:培养小鼠RAW264.7巨噬细胞,采用IL-4诱导并建立M2型极化模型,将培养细胞分为未诱导细胞(M0)组、M2极化(IL-4诱导)组、MPRR诱导组和Rp组(IL-4诱导12 h后MPRR再极化诱导组),流式细胞术分析巨噬细胞极化标志(CD86和CD206)的表达。免疫印迹技术分析转录因子STAT-6表达和磷酸化。收集不同条件下巨噬细胞培养上清液,ELISA法检测其IFN-γ和TGF-β浓度。结果:与对照组比较,IL-4诱导后细胞发生M2极化,可检测到RAW264.7细胞表面CD86表达下降(P0.05),CD206表达增加(P0.05)。而MPRR促进细胞向M1极化,可观察到其促进CD86表达而降低CD206表达(P0.05)。M2极化后STAT-6磷酸化增加,IFN-γ分泌减少而TGF-β分泌增加(P0.05);经MPRR处理减弱STAT-6磷酸化和TGF-β的分泌,促进IFN-γ分泌(P0.05)。结论:炙甘草水溶性多糖促进巨噬细胞的M1极化,可拮抗IL-4诱导其M2极化效应,炙甘草多糖可调节小鼠巨噬细胞再极化。     

N-Ac-PGP通过激活TLR4诱导巨噬细胞的M1极化并提高慢性阻塞性肺疾病炎症水平

目的：探究N-乙酰化脯氨酸-甘氨酸-脯氨酸（N-acetylated proline-glycine-proline, N-Ac-PGP）通过Toll样受体4(Toll-like receptor 4, TLR4)诱导巨噬细胞M1极化对慢性阻塞性肺疾病（chronic obstructive pulmonary disease, COPD）炎症反应的影响。方法：检测COPD患者痰液中N-Ac-PGP和M1型巨噬细胞炎性细胞因子的表达。体外培养人单核细胞白血病细胞THP-1,使用佛波脂（phorbol 12-myristate 13-acetate, PMA）诱导使之分化为M0型巨噬细胞,然后使用脂多糖（lipopolysaccharide, LPS）和N-Ac-PGP诱导M0型巨噬细胞极化为M1型。采用RTqPCR和Western blot法分别检测巨噬细胞表面标志物CD11b、CD45、CD86和诱导型一氧化氮合酶（inducible nitric oxide synthase, iNOS）的mRNA和蛋白表达水平。使用ELISA检测M1型巨噬细胞炎性细胞因子肿瘤坏死因子α(tu-m...     

Activated phenotype in neutrophils and monocytes from patients with primary proliferative polycythaemia.

AIM--To investigate whether monocytes and neutrophils from patients with primary proliferative polycythaemia (PPP) exhibit increased expression of markers of cell activation and, if so, whether they are associated with the phagocytic activity of these cells and concentrations of circulating cytokines. METHODS--Expression of CD11b, CD14, CD18, and CD64 on monocytes and neutrophils was assessed by flow cytometry. Phagocytosis was analysed using immunoglobulin opsonised Escherichia coli. Serum concentrations of granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF) and macrophage CSF (M-CSF) were determined by bioassays, and interferon-gamma (IFN-gamma) by enzyme linked immunosorbent assay (ELISA). RESULTS--Patients with PPP (n = 18), when compared with normal subjects (n = 10), had increased percentages of CD64+ monocytes (52% v 36%) and neutrophils (42% v 11%) and of CD14+ neutrophils (36% v 18%). Monocytes from patients with PPP exhibited increased expression of CD64 (47 v 26) and of CD11b (65 v 36). These abnormalities were not found in patients with secondary (n = 8) or apparent (n = 13) polycythaemia. The percentage of neutrophils undergoing phagocytosis was higher in patients with PPP (mean 64%; n = 6) than in normal subjects (mean 42%; n = 5). G-CSF, GM-CSF and IFN-gamma concentrations in patients' serum samples were comparable with normal; M-CSF was not detected in any of the samples. There was no correlation between cytokine concentrations and the expression of CD11b, CD14, CD18, and CD64 on patients' phagocytes. CONCLUSIONS--Increased expression of CD11b and CD64 by monocytes, increased percentages of CD14+ and CD64+ neutrophils and the high phagocytic activity of neutrophils suggests that these cells are activated in vivo in patients with PPP. The phenotypic changes of PPP phagocytes were not associated with increased concentrations of circulating cytokines and probably reflect intrinsic abnormalities within the neoplastic PPP clone.     

IL-10 augments CD23 expression on U937 cells and down-regulates IL-4-driven CD23 expression on cultured human blood monocytes: effects of IL-10 and other cytokines on cell phenotype and phagocytosis.

The effects of human recombinant interleukin-10 (IL-14) on the expression of several markers on U937 and human peripheral blood monocytes was studied by immunofluorescence and fluorescence-activated cell sorter (FACS) analysis. IL-10 augmented Fc IgE receptor (Fc epsilon RII/CD23) further enhanced by cotreatment with IL-4 or interferon-gamma (IFN-gamma). In contrast, the basal level of Fc epsilon RII expression on blood monocytes appeared to fall in response to IL-10, and this effect became more evident on IL-4-treated cells. Furthermore, the constitutive and IFN-gamma-triggered Fc gamma RI/CD64 expression was augmented on both monocytes and U937 cells. Thus the expression of Fc gamma RII/CD32, Fc gamma/RIII/CD16, Fc alpha R/CD89, the receptor for complement components (CR1/CD35, CD3/CD11b, CR4/CD11c) and the receptor for transferrin/CD71 was not significantly influenced on IL-10-treated cells. IL-10 modestly triggered CD14 antigen expression on monocytes but not U937. The expression of intercellular adhesion molecule-1 (ICAM-1)/CD54 on monocytes was significantly inhibited by IL-10. As expected, a marked reduction of the constitutive as well as of the IFN-gamma or IL-4-driven expression on HLA-DR, HLA-DP and HLA-DQ was observed on IL-10-cultured monocytes. On the other hand, the expression of major histocompatibility complex (MHC) class I molecules was slightly and dose-dependently induced on IL-10-treated monocytes. The ability of blood monocytes to phagocytose IgG-sensitized ox erythrocytes, and to bind and ingest opsonized Escherichia coli or latex particles, was amplified by IL-10. Our data demonstrate that IL-10 modulates the expression of a wide variety of structures on human mononuclear phagocytes, and augments their phagocytic capacity.     

Membrane molecules which trigger the production of interleukin-1 and tumor necrosis factor-alpha by lipopolysaccharide-stimulated human monocytes.

We have investigated the role of the membrane molecules CD11/CD18 and CD14 which may mediate the binding of lipopolysaccharide (LPS) to human monocytes, in the induction of the production and release of interleukin (IL)-1 and tumor necrosis factor-alpha (TNF-alpha) by LPS-stimulated cells. Blockade of CD11a, CD11b and CD18 with saturating concentrations of specific mAb did not inhibit the release of cytokines from LPS-stimulated monocytes. In contrast, inhibition of the release of IL-1 beta and TNF-alpha occurred in monocytes cultures that had been pretreated with either of two monoclonal antibodies (mAb) recognizing different epitopes on the CD14 molecule. The binding of LPS to CD14 has been previously shown to require serum factors. In the present study, we found that serum had an enhancing effect on the release of IL-1 and TNF-alpha from LPS-stimulated cultures of normal human monocytes. The inhibitory effect of anti-CD14 mAb was, however, observed in cultures performed in the presence or in the absence of serum, suggesting that triggering of IL-1/TNF-alpha release by CD14 is independent of LPS-binding proteins or other serum proteins. IL-1 beta and TNF-alpha were also released from LPS-stimulated cultures of monocytes from patients with paroxysmal nocturnal hemoglobinuria lacking expression of CD14. Thus, CD14 but not CD11/CD18 can trigger serum-dependent and independent cytokine release from endotoxin-stimulated normal human monocytes; CD14 is not, however, the only LPS receptor that is involved in the secretory response of endotoxin-stimulated cells.     

Expansion and differentiation of CD14+CD16(-) and CD14+ +CD16+ human monocyte subsets from cord blood CD34+ hematopoietic progenitors

To determine whether monocytes can be generated from CD34+ hematopoietic progenitors in large numbers, cord blood CD34+ cells were first expanded for 3-10 days in X-VIVO 10 medium supplemented with FCS, stem cell factor (SCF), thrombopoietin (TPO), and Flt-3 Ligand (Flt-3L), and then differentiated in IMDM medium supplemented with FCS, SCF, Flt-3L, IL-3 and M-CSF for 7-14 days. These two step cultures resulted in up to a 600-fold mean increase of total CD14+ cells. Using this approach, two subpopulations of monocytes were obtained: CD14+CD16(-) and CD14++CD16+ occurring at 2:1 ratio. 1.25(OH)2 Vitamin D3 added to the differentiation medium altered this ratio by decreasing proportion of CD14++CD16+ monocytes. In comparison to CD14+CD16(-), the CD14++CD16+ cells showed different morphology and an enhanced expression of CD11b, CD33, CD40, CD64, CD86, CD163, HLA-DR, and CCR5. Both subpopulations secreted TNF and IL-12p40 but little or no IL-10. CD14++CD16+ monocytes released significantly more IL-12p40, were better stimulators of MLR but showed less S. aureus phagocytosis. These subpopulations are clearly different from those present in the blood and may be novel monocyte subsets that represent different stages in monocyte differentiation with distinct biological function.     

HIV-1 does not alter in vitro and in vivo IL-10 production by human monocytes and macrophages

The present study analyses the ability of HIV-1 to modulate IL-10 production in cells of monocyte-macrophage lineage cultured in the presence of macrophage colony-stimulating factor (M-CSF). Both monocytes and macrophages spontaneously produced low amount of IL-10. Lipopolysaccharide (LPS) induced a strong IL-10 response in fresh monocytes and in M-CSF-treated macrophages. In contrast, macrophages cultured in the absence of M-CSF exhibited a marked decrease in their susceptibility to LPS stimulation. M-CSF increased the IL-10 response of macrophages to LPS by enhancing both the expression of membrane-bound CD14, the protein that serves as LPS receptor, and the sensibility of CD14-expressing cells to LPS stimulation. Neither spontaneous nor LPS-induced expression of IL-10 was modulated in monocytes and macrophages by infection with eight monocytotropic strains, as demonstrated by ELISA and cytofluorimetric analysis. In contrast, all the HIV-1 strains primed macrophages for an increased IL-6 response to LPS stimulation. To determine whether IL-10 production was associated with in vivo infection, monocytes from AIDS individuals were analysed for IL-10 production. We found that neither spontaneous nor LPS-induced IL-10 production were different between healthy controls and HIV-infected patients. Taken together, these data strongly suggest that HIV-1 infection of monocytes-macrophages does not play a significant role in the regulation of IL-10 in infected patients. This study also emphasizes the role of M-CSF activation in the regulation of the cytokine response in macrophages.     

HIV-1 induces IL-10 production in human monocytes via a CD4-independent pathway

In HIV-infected patients, increased levels of IL-10, mainly produced by virally infected monocytes, were reported to be associated with impaired cell-mediated immune responses. In this study, we investigated how HIV-1 induces IL-10 production in human monocytes. We found that CD14(+) monocytes infected by either HIV-1(213) (X4) or HIV-1(BaL) (R5) produced IL-10, IL-6, tumor necrosis factor-alpha (TNF-alpha), and to a lesser extent, IFN-gamma. However, the capacity of HIV-1 to induce these cytokines was not dependent on virus replication since UV-inactivated HIV-1 induced similar levels of these cytokines. In addition, soluble HIV-1 gp160 could induce CD14(+) monocytes to produce IL-10 but at lower levels. Cross-linking CD4 molecules (XLCD4) with anti-CD4 mAbs and goat anti-mouse IgG (GAM) resulted in high levels of IL-6, TNF-alpha and IFN-gamma but no IL-10 production by CD14(+) monocytes. Interestingly, neither anti-CD4 mAbs nor recombinant soluble CD4 (sCD4) receptor could block IL-10 secretion induced by HIV-1(213), HIV-1(BaL) or HIV-1 gp160 in CD14(+) monocytes, whereas anti-CD4 mAb or sCD4 almost completely blocked the secretion of the other cytokines. Furthermore, HIV-1(213) could induce IL-10 mRNA expression in CD14(+) monocytes while XLCD4 by anti-CD4 mAb and GAM failed to do so. As with IL-10 protein levels, HIV-1(213)-induced IL-10 mRNA expression in CD14(+) monocytes could not be inhibited by anti-CD4 mAb or sCD4. Taken together, HIV-1 binding to CD14(+) monocytes can induce CD4-independent IL-10 production at both mRNA and protein levels. This finding suggests that HIV induces the immunosuppressive IL-10 production in monocytes and is not dependent on CD4 molecules and that interference with HIV entry through CD4 molecules may have no impact on counteracting the effects of IL-10 during HIV infection.     

Interleukin-3, -5, and granulocyte macrophage colony-stimulating factor-induced adhesion molecule expression on eosinophils by p38 mitogen-activated protein kinase and nuclear factor-[kappa] B

We investigated the intracellular signaling mechanisms for cytokine interleukin (IL)-3, IL-5, or granulocyte-macrophage colony-stimulating factor (GM-CSF)-induced expression of adhesion molecules including very late antigen 4 (CD49 d), macrophage antigen-1 (CD11b), leukocyte function-associated antigen-1 (CD11a/CD18), intercellular adhesion molecule (ICAM)-1, and ICAM-3 on eosinophils. The expression of adhesion molecules and nuclear factor (NF)-kappaB pathway was measured by flow cytometry and cDNA expression array, respectively. The phosphorylation of inhibitor kappaB-alpha and p38 mitogen-activated protein kinase (MAPK) was detected by Western blot, whereas NF-kappaB activity was measured by electrophoretic mobility shift assay. IL-3, IL-5, and GM-CSF could enhance p38 MAPK and NF-kappaB activity and induce ICAM-1, CD11b, and CD18 expressions on eosinophils. They could suppress ICAM-3 expression, but had no effect on CD49 d expression. Either SB 203580 or MG-132 was able to offset the cytokine-induced expression of ICAM-1. Only SB 203580 could reverse the effect on CD11b, CD18, and ICAM-3 expressions. Therefore, the expression of ICAM-1 might involve both p38 MAPK and NF-kappaB activities, whereas the regulation of CD11b, CD18, and ICAM-3 expressions might be mediated through p38 MAPK but not NF-kappaB. These cytokines therefore play a crucial role, via the p38 MAPK and NF-kappaB pathways, in the expression of important adhesion molecules on eosinophils in allergic inflammation.     

Changed phagocytic activity and pattern of Fcγ and complement receptors on blood monocytes in sarcoidosis

We have recently revealed that mycobacterial heat shock proteins (Mtb-hsp), involved in forming of immune complexes (CIs), can induce immune response in sarcoidosis (SA). The complexemia may result from inappropriate phagocytosis and clearance of CIs by monocytes with following persistent antigenemia and granuloma formation. Because an aberrant expression of receptors for Fc fragment of immunoglobulin G (FcγR) and complement receptors (CR) on monocytes can be involved in this process, we have evaluated the expression of FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16) and CR1 (CD35), CR3 (CD11b), CR4 (CD11c) receptors on blood CD14(+) monocytes and its phagocytic activity in 24 patients with SA and 20 healthy volunteers using flow cytometry. We found significantly increased expression of all examined FcγR and decreased expression of CD35 and CD11c on CD14(+) monocytes in SA patients vs controls. Significantly increased percentage of CD14(+)CD16(+)CD35(-), CD14(+)CD64(+)CD35(+), CD14(+)CD64(+)CD11b(+), CD14(+)CD64(+)CD11c(+) and decreased of CD14(+)CD32(-)CD35(+), CD14(+)CD32(-)CD11b(+), CD14(+)CD32(-)CD11c(+) monocytes' phenotypes was revealed in SA. The total number and percentage of phagocyting monocytes was significantly increased in SA as compared with controls. In conclusion, altered expression of FcγR and CR on CD14(+) monocytes and its increased phagocytic activity may be responsible for high antigen load, persistent antigenemia and immunocomplexemia in SA patients.