Blood Lymphocytes, Monocytes and NK Cells Modulate Their Expression of CD44, ICAM-1, LFA-1 and MHC Class II After Arrival Into Lymphoid Organs

Adhesion molecules expressed on surface membranes of lymphocytes and other leukocytes enable their entry into the lymphoid and other tissues. However, little is known about molecules that govern the transit of leukocytes through the parenchyma of lymphoid organs proper. We show that in comparison to blood leukocytes, the corresponding cells isolated from lymphoid organs, i.e., lymph nodes and spleen, have a significantly augmented expression of certain surface molecules. The helper and cytotoxic subsets of T cells, as well as B cells, display the increased expression of CD44, ICAM-1 and LFA-1, whereas B cells additionally show the augmented expression of MHC class II. In comparison with blood monocytes, splenic monocytes show the increased expression of ICAM-1 and MHC class II molecules. When compared with blood NK cells, splenic NK cells only show the increased expression of ICAM-1. The molecules, which we show to be up regulated upon the entry of leukocytes into lymphoid organs, could be involved in their retention within the tissue via cell-cell or cell-extracellular matrix interactions and in control of their transit through lymphoid tissues.     

人外周血淋巴细胞及单核细胞表达PD-L1和PD-L2的动力学研究

目的： 探讨PD-L1和PD-L2在正常人外周血静息和活化的B细胞、T细胞及单核细胞表面的表达规律。 方法： 利用荧光抗体染色和流式细胞术分析静息状态及经多克隆刺激剂脂多糖（LPS）和美洲商陆丝裂原（PWM）刺激6、24、48和72 h后表达PD-L1和PD-L2的B细胞和T细胞百分率，同时分析静息状态及经IFN-γ和LPS共同刺激24、48、72 和96 h后表达PD-L2的单核细胞百分率。 结果： 静息B细胞和T细胞表面不表达PD-L1，LPS和PWM刺激6 h后表达PD-L1的B细胞百分率均显著高于静息B细胞，24 h达到最高，分别为(46.26±10.71)%和(43.67±6.14)%，之后随时间延长而降低；表达PD-L1的T细胞百分率在LPS刺激下无显著变化，但PWM刺激6 h后百分率明显高于静息条件，24 h达到最高，为(25.42±9.23)%，之后下降。静息B细胞和T细胞不表达PD-L2，活化后PD-L2表达也无明显上调。另外，静息状态单核细胞表面不表达PD-L2，体外活化24 h后表达PD-L2的单核细胞百分率显著上调，48 h达到最高为(28.70±14.22)%，之后随时间而降低。 结论： 活化的淋巴细胞表达PD-L1，但不表达PD-L2，后者在活化的单核细胞表面表达，并且后者达到高峰的时间迟于前者。     

Role of T lymphocytes in the pathogenesis of experimental autoimmune encephalomyelitis

A dichotomy between clinical and histological signs of experimental autoimmune encephalomyelitis (EAE) was demonstrated in neonatally thymectomized (NNTx) rats. Although neonatal thymectomy prevented subsequent induction of clinical EAE in Lewis rats challenged as young adults with syngeneic basic protein (BP) of myelin, there was a 50 % incidence of histological EAE. Both cellular and humoral immune responses to BP were reduced in the NNTx rats. However, the presence of a cell-mediated response to BP (MIF release) was statistically associated with the occurrence of histological EAE. Diminished preimmunization blood lymphocyte counts and impairment of the response of spleen cells to phytohemagglutinin were associated with reduced antibody responses. This suggests that collaboration between T and B cells may be necessary for production of antibody to BP. The relationship between antibody and clinical EAE is not clear and warrants further investigation.     

Granulocytes as active participants in acute myocardial ischemia and infarction

The classical picture of the polymorphonuclear leukocyte--antimicrobal activities and participation in the inflammatory reaction--appears now to be expanded. It has been recognized that this cell may be highly injurious to other cells, has a propensity to be trapped in the capillary network, and may occlude microvascular pathways. The classical hypothesis that the granulocytes appear in the ischemic or postischemic myocardium as response to the associated inflammatory reaction may have to be revised. Instead, the granulocyte may actually be the key factor causing the inflammation. An array of future studies is needed to clarify the events step by step. Important aspects are the activation and degranulation sequence in the ischemic myocardium, the form and extent of injury caused by the oxygen free radicals and lysosomal enzymes, the source and nature of chemotactic substances, particularly in human forms of myocardial ischemia, and the degree to which granulocytes cause or magnify the injury.     

Menopause Is Associated With a Significant Increase in Blood Monocyte Number and a Relative Decrease in the Expression of Estrogen Receptors in Human Peripheral Monocytes

PROBLEM : The clinical significance of the differential expression of estrogen receptor (ER) in human monocytes was evaluated. METHOD : Two color flow cytometry analysis was used on peripheral blood samples of young and postmenopausal females and postmenopausal females treated with estrogen replacement therapy. In addition, the monocyte and lymphocyte counts and the blood estrogen levels of each patient were determined. RESULTS : During menopause there is a significant decrease in the percentage of ER positive monocytes, and an increase in blood monocyte number, which declines following estrogen replacement therapy to values of the young. CONCLUSIONS : These findings suggest that estrogen modulates the monocyte numbers and its effects may be mediated through the ER in the monocytes.     

CXCR4 and CCR5 ligands cooperate in monocyte and lymphocyte migration and in inhibition of dual-tropic (R5/X4) HIV-1 infection

One of the most important functions of chemokines and their receptors is the regulation of directional migration of leukocytes within tissues. In specific tissue compartments, cells are exposed to multiple chemokines presented in complex dimensional and temporal patterns. Therefore, a leukocyte requires the mechanisms to integrate the various directional signals it receives from different chemoattractants. In this study, we report that CCL3, CCL5, and CCL8, three potent mononuclear cell chemoattractants, are able to synergize with the homeostatic chemokine CXCL12 in the migration of CD14(+) monocytes, CD3(+) T-lymphocytes, or PHA-activated lymphoblasts. In addition, CCL5 augmented the CXCR4 ligand-driven ERK phosphorylation in mononuclear cells. Furthermore, the synergistic effect between CCL5 and CXCL12 in monocyte chemotaxis is inhibited in the presence of specific CCR1 antibody and AMD3100, but not by maraviroc. In HIV-1 infection assays, a combination of CXCL12 and CCL5 cooperated to inhibit the replication of the dual-tropic (R5/X4) HIV-1 HE strain. Finally, although the dual-tropic HIV-1 strain was barely suppressed by AMD3100 or maraviroc alone, HIV-1 infection was completely blocked by the combination of these two receptor antagonists. Our data demonstrate the cooperation between CCL5 and CXCL12, which has implications in migration of monocytes/lymphocytes during inflammation and in HIV-1 infection.     

Intraepithelial lymphocytes and macrophages in the normal breast

Summary In this study the presence of intraepithelial cells within the normal breast parenchyma was investigated by electron microscopy and immunocytochemistry. Cells were observed which could be differentiated from the epithelial and myoepithelial cells by their cytoplasmic and nuclear morphology and the absence of cell junctions. Two cell types (lymphocytes and macrophages) were identified ultrastructurally and the bone marrow origin of the cells was confirmed by immunocytochemistry. The intraepithelial lymphocytes and macrophages were present in all samples irrespective of the physiological state. In the resting, pregnant, and lactating breast the majority of cells were lymphocytes while in the involuting breast there was a marked increase in the proportion of macrophages. The rarity of lymphoma of the breast may be related to the relatively small amount of lymphoid tissue present and the passive nature of the environment.     

Monocytes as effector cells: activated Ly-6C(high) mouse monocytes migrate to the lymph nodes through the lymph and cross-present antigens to CD8+ T cells

Monocytes have the capacity to differentiate into macrophages or dendritic cells (DCs) after extravasation into lymphoid and nonlymphoid tissues. They have thus been consequently considered as precursors, but not effector cells, recirculating exclusively through the blood. In this report, we demonstrate for the first time that, after subcutaneous injection, activated monocytes migrate through the lymphatics from the dermis into the draining lymph nodes by a CCR7-dependent mechanism. LPS-activated monocytes were less efficient than DCs in stimulating CD4(+) T cells, but unexpectedly, they were highly efficient in inducing antigen-specific CD8(+) T-cell proliferation by cross-presentation, both in vitro and in vivo. Interestingly, CD8(+) T cells stimulated in vivo by activated monocytes expressed a high level of CD62L, suggesting that they had undergone an unconventional activation process. In conclusion, our data strongly support the concept that monocytes can behave not only as precursor cells for macrophages and DCs, but also as effector cells with the capacity to migrate from the periphery to the lymph nodes through the lymph and to cross-present antigens to CD8(+) T cells. These results suggest that monocytes can play an important role in the induction and regulation of CD4(+) and CD8(+) T-cell responses.     

Peculiarity of small lymphocytes in the thymic medulla in neonatal mice

Summary Karyometric studies were made of lymphocytes for the cortex and medulla of the thymus of mice at various ages from birth to adulthood. From the results obtained it was evident that in early neonate mice, medullary small lymphocytes were characterized by having larger nuclei than cortical small lymphocytes. On the basis of cytological features, such medullary lymphocytes could reasonably be classified as small lymphocytes rather than medium lymphocytes, although they had larger nuclei than typical small lymphocytes. Such peculiar small lymphocytes with larger nuclei were preponderant in the medulla during early neonatal life, but they rapidly decreased in number with advancing age, although they were present in a small proportion even in adults. The findings on the occurrence of such peculiar small lymphocytes in the medulla were discussed in relation to the maturation pathway of lymphocytes in the thymus.Supported by research grant from the Education Ministry (No. 748046, 1972).     

茴香霉素在小鼠淋巴细胞增殖和活化中的作用

目的研究茴香霉素在小鼠淋巴细胞增殖和活化中的作用。方法以活体染料羧基荧光素乙酰乙酸琥珀酰亚胺酯染色，建立了在多克隆刺激剂刀豆蛋白A（ConA）刺激下评价小鼠淋巴细胞增殖的模型，通过流式细胞术及MTT法分析茴香霉素在不同剂量下对淋巴细胞增殖的作用；采用碘化丙锭染色分析茴香霉素对ConA或佛波醇酯（PDB）加离子霉素（Ion）刺激的小鼠淋巴细胞周期变化的作用；利用荧光标记的单克隆抗体双染技术结合流式细胞仪检测茴香霉素对小鼠CD3^＋淋巴细胞早期及中期活化标志分子CD69和CIY25表达的影响。结果随着茴香霉素浓度从1．0ng／ml逐渐增至25．0ng，ml，ConA对淋巴细胞的促增殖作用逐渐减弱，以10．0ng，ml和25．0ng，ml茴香霉素的抑制作用最为明显，呈剂量依赖关系（r=-0．96，P〈0．01）。进一步发现，1．0—25．0ng/ml茴香霉素能够使ConA或PDB加Ion诱导的淋巴细胞停滞于G0/G1期，阻止其进入S期，且阻止作用随上述浓度的增加而增强，呈明显剂量依赖关系（r=-0．97，P〈0．01和r=-0．98，P〈0．01）。据此选用最佳剂量10．0ng/ml，茴香霉素能够明显抑制淋巴细胞表面分子CD69和CIY25的表达（P均〈0．01）。结论茴香霉素能明显抑制小鼠淋巴细胞的增殖和活化。     

Carvedilol Modulates In-Vitro Granulocyte-Macrophage Colony-Stimulating Factor-Induced Interleukin-10 Production in U937 Cells and Human Monocytes

Both granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-10 (IL-10) are important mediators regulating inflammatory responses. Inflammatory processes have an important role in atherogenesis. In this paper, the effects of carvedilol on GM-CSF-induced IL-10 production were examined on human monocytic cell line, U937, and purified human monocytes. First, we showed that one-time carvedilol pretreatment at concentrations 0.3-10 μM dose-dependently inhibited GM-CSF-induced IL-10 production in U937 cells. In addition, we found carvedilol to be non-cytotoxic at concentrations equal to or less than 10 μM. However, at concentrations higher than 10 μM, carvedilol induced programmed cell death in U937 cells. The inhibition of GM-CSF-induced IL-10 production by carvedilol was also observed at the expression of mRNA. Furthermore, the inhibition of IL-10 production was demonstrated in GM-CSF-activated purified human peripheral blood monocytes. Finally, long-term carvedilol pretreatment of U937 cells up to 2 months at concentrations of 1.0 μM mildly enhanced the IL-10 production. Our observations that carvedilol modulated GM-CSF-induced IL-10 production may have some implication in understanding the broad-spectrum effects of carvedilol in regulating inflammatory reactions.     

杂色曲霉素对体外培养HPBMcIFN-γ分泌的影响

目的:探讨杂色曲霉素(ST)对人T淋巴细胞分泌功能的影响。方法:采用双抗体夹心ELISA方法对ST作用后人外周血单核细胞(HPBMc)培养上清液中γ-干扰素(IFN-γ)的分泌水平进行检测。结果:不同浓度ST对IFN-γ分泌的影响不同,较低浓度ST0.03125mg/L-0.12500mg/L对IFN-γ的分泌有一定抑制作用(P＜0.05),而在0.25mg/L以上浓度ST可刺激IFN-γ分泌,至1mg/L达到高峰(P＜0.05)。在0.25mg/L-1.00mg/L的浓度范围内,ST浓度和IFN-γ分泌呈正相关(r=0.492,P＜0.01);在ST1mg/L作用1h-64h的时间范围内,ST对IFN-γ分泌的影响依时间不同作用也不同,在4h-8hIFN-γ的分泌降低(8h,P＜0.05),而以后各组随着ST作用时间的延长IFN-γ水平逐渐增高,尤以32h组增高最为明显(P＜0.05),在ST作用16h-64h的时间范围内,IFN-γ的分泌随着ST作用时间的延长而逐渐增加,两者呈正相关(r=0.736,P＜0.01)。结论:ST对HPBMcIFN-γ分泌的影响表现在较低浓度和较短时间抑制IFN-γ分泌,而较高浓度和较长时间则诱导IFN-γ分泌,但继续增加ST浓度和作用时间并不继续增加IFN-γ分泌。     

肿瘤浸润性淋巴细胞和肿瘤坏死因子α协同抗肿瘤作用的研究

利用体外消化结合不连续密度梯度离心的方法，由肿瘤组织内分离出肿瘤浸润性淋巴细胞（ＴＩＬ），并在重组人类白细胞介素２（ｒＩＬ－２）和／或重组肿瘤坏死因子α（ｒＴＮＦα）存在下进行体外培养。结果表明，低浓度ｒＴＮＦａ可以促进ＴＩＬ体外增殖、增加处于Ｓ期及Ｇ＿２＋Ｍ期的ＴＩＬ比例、增加细胞毒Ｔ细胞比例、上调ＴＩＬＩＬ－２Ｒ表达，并能明显增强ＴＩＬ体内外对自体及同种异体肿瘤的杀伤活性。     

黄曲霉毒素G1对体外培养HPBM增殖及TNF-α分泌的影响

目的:研究黄曲霉毒素G₁(AFG₁)对体外培养人外周血单个核细胞(HPBM)增殖及细胞TNF-α分泌的影响。方法:采用流式细胞术(FCM)和MTT比色法研究AFG₁对HPBM增殖的影响。以双抗体夹心ELISA法检测AFG₁对HPBMTNF-α分泌的影响。结果:FCM检测结果显示,AFG₁作用6h,1000μg/L处理组HPBM的增殖指数明显高于对照组。AFG₁作用24h,200μg/L和1000μg/L浓度的AFG₁可明显刺激HPBM增殖;回归分析结果表明,AFG₁作用6h和24h,AFG₁浓度均与增殖指数呈正相关(r分别为0.5122和0.5119,P均＜0.05)。MTT比色法结果显示,2000μg/LAFG₁处理HPBM的A值明显高于对照组。AFG₁在100μg/L浓度下可显著抑制TNF-α分泌(P＜0.05)。结论:AFG₁对体外培养HPBM的增殖有刺激作用,在100μg/L浓度下对HPBMTNF-α的分泌有一定抑制作用。     

Differentiation of lymphocytes in cerebrospinal fluid in disorders of the central nervous system

Summary In 73 patients with variable CNS-disorders Slg-lymphocytes and RF-lymphocytes originating from CF were characterized by demonstration of surface markers. In the CF the Slg-lymphocytes were increased in comparison to the blood whereas the RF-lymphocytes generelly were decreased. Patients suffering from acute or chronic inflammatory diseases had significantly more Slg-lymphocytes while RF-lymphocytes were significantly decreased in those patients as well as in patients with malignancies. A correlation between the number of Slg-lymphocytes and immunoglobulin-levels was not seen. The respective data in blood did not follow the changes in CF. The results suggest a prevailing stimulation of the humoral immune system of the CNS during inflammatory diseases.