Up-regulation of Toll-like receptors 2, 3 and 4 in allergic rhinitis

Background

Toll-like receptors enable the host to recognize a large number of pathogen-associated molecular patterns such as bacterial lipopolysaccharide, viral RNA, CpG-containing DNA and flagellin. Toll-like receptors have also been shown to play a pivotal role in both innate and adaptive immune responses. The role of Toll-like receptors as a primary part of our microbe defense system has been shown in several studies, but their possible function as mediators in allergy and asthma remains to be established. The present study was designed to examine the expression of Toll-like receptors 2, 3 and 4 in the nasal mucosa of patients with intermittent allergic rhinitis, focusing on changes induced by exposure to pollen.

Methods

27 healthy controls and 42 patients with seasonal allergic rhinitis volunteered for the study. Nasal biopsies were obtained before and during pollen season as well as before and after allergen challenge. The seasonal material was used for mRNA quantification of Toll-like receptors 2, 3 and 4 with real-time polymerase chain reaction, whereas specimens achieved in conjunction with allergen challenge were used for immunohistochemical localization and quantification of corresponding proteins.

Results

mRNA and protein representing Toll-like receptors 2, 3 and 4 could be demonstrated in all specimens. An increase in protein expression for all three receptors could be seen following allergen challenge, whereas a significant increase of mRNA only could be obtained for Toll-like receptor 3 during pollen season.

Conclusion

The up-regulation of Toll-like receptors 2, 3 and 4 in the nasal mucosa of patients with symptomatic allergic rhinitis supports the idea of a role for Toll-like receptors in allergic airway inflammation.     

变应性鼻炎患者外周血中IL-27和Treg细胞的相关性分析

目的探讨外周血IL-27和CD4^＋CD25^＋调节性T细胞（Treg）在变应性鼻炎（AR）发病机制中的作用。方法2012年3月至7月,收集AR患者32例（AR组）和20例健康志愿者（对照组）外周血,采用流式细胞术（FCM）检测外周血中Treg细胞比例;ELISA检测血清中IL-27、IL-10和TGF-β1的水平。结果AR组Treg细胞百分率[（1．75±0．56）％]明显低于对照组[（4．76±1．75）％],两组比较的差异有统计学意义（P〈0．01）。AR组IL-27、IL-10和TGF-β1的水平分别为（24．43±16．36）pg／ml、（14．29±6．16）pg／ml、（0．34±0．04）pg／ml,均低于对照组（44．09±13．12）pg／ml、（31．32±21．20）pg／ml、（O．49±0．06）pg／ml,两组之间的差异有统计学意义（P〈0．01）。AR患者外周血中IL-27和Treg细胞百分率、IL-10、TGF-β1存在正相关（r分别为0．825,0．646,0．517,P〈0．01）,Treg细胞百分率和IL-10、TGF-β1存在正相关（r=0.622,0．738,P〈0．01）,IL-10和TGF-β1无相关性（r=0．304,P〉0．05）结论AR患者外周血中IL-27水平降低,Treg细匏百分率降低及其主要分泌因子IL-10、TGF-β1水平降低,且IL-27与Treg细胞百分率、IL-10、TGF-β1水平呈正相关,提示在AR发病中IL-27对Treg细胞可能具有免疫调节作用。     

CFU-gm colony formation of peripheral blood and bone marrow in adult acute leukemia at presentation, during remission, and at relapse

Granulocyte/macrophage colony-forming unit (CFU-gm) formation was studied simultaneously in bone marrow and peripheral blood of 52 previously untreated adult patients with acute non-lymphocytic (ANLL) and 36 with acute lymphoblastic leukemia (ALL). They were followed during induction therapy at monthly intervals while in remission and in 19 ANLL and 22 ALL cases, until relapse. Patients showing a decreased colony number in the marrow but normal or increased colony numbers in the peripheral blood had a high probability of entering remission. Non-responding patients displayed an opposite pattern. The higher the degree of marrow repopulation with granulocytic progenitor cells after induction treatment, the longer remission duration and survival for ANLL patients and the longer survival for ALL patients. CFU-gm formation returned to normal in the early stages of complete remission, but then declined progressively. At ANLL and ALL relapse, colony growth was reduced markedly while cluster formation remained normal. The number of marrow colonies and clusters in ANLL were significantly higher at first and second relapse compared to the growth pattern at first presentation. A similar trend had been observed in ALL, suggesting a selection advantage.     

The time-course of micronucleated polychromatic erythrocytes in mouse bone marrow and peripheral blood

The time-course of micronucleated polychromatic erythrocytes (MPCE) in mouse bone marrow and peripheral blood, induced by an acute 0.1 Gy dose of X-rays, was determined using flow cytometric analysis, which made frequent sampling possible and allowed use of a dose low enough not to affect erythroid cell proliferation. The frequency of MPCE (fMPCE) began to increase in the bone marrow at 10 h after irradiation and reached a maximum at 28 h after irradiation. In the peripheral blood fMPCE began to increase at 20 h after irradiation and peaked at about 40 h after irradiation. The time-course found is discussed on the basis of data on the differentiation of erythroid cells. The results indicate that the micronuclei registered in polychromatic erythrocytes may originate from lesions induced not only during the last cell cycle but also during earlier ones. After an acute dose of 1.0 Gy of X-rays the maximum fMPCE was delayed both in bone marrow and peripheral blood reflecting an effect on the cell cycle progression of erythroblasts.     

Changes of bone marrow cell and peripheral blood cell counts in newborn mice

Differential bone marrow cell and peripheral blood cell counts in new-born mice from zero to twenty-one days of age were investigated. In the bone marrow, erythroblasts rapidly increased whereas granulocytic cells decreased with time. The G/E ratio was 6.7 on day zero and 0.5 on day four, and as the G/E ratio of adult mice is normally from 0.8 to 2.4, these values would be the highest and lowest respectively for the whole life span of a mouse. In the peripheral blood, MCV decreased each day, closely matching the changes in the Price-Jones curves. The results of erythrocyte area measurements indicate that the larger the erythrocyte area, the earlier in days of age it disappeared and the shorter its life span.     

Lymphocyte subpopulations in peripheral blood and bone marrow in patients with idiopathic myelofibrosis

In 8 patients with idiopathic myelofibrosis (IM) T and B cells were studied in view of the possibility that immunological dysfunctions are involved in initiating or contributing to the bone marrow fibrosis. In peripheral blood the absolute numbers of E-SRBC and OKT3+ lymphocytes were significantly reduced; in addition a significant decline was observed in the proportion and absolute numbers of OKT8+ cells, resulting in a reversed Leu-3a/OKT8 ratio. An impaired B cell function was observed in 4 of the 8 patients, characterized by a disturbed in vitro pokeweed mitogen stimulated immunoglobulin synthesis and low serum immunoglobulin levels. Immuno-histological studies of the bone marrow demonstrated a scarcity of T cells but normal numbers of B cells. However, no correlation was noted between the observed deviations of B and T cells and the degree of bone marrow fibrosis determined by means of bone marrow histology and serum procollagen-III levels. These data are not sufficient to support the hypothesis that immunological changes in IM are primarily involved in the process of bone marrow fibrosis.     

小鼠Toll样受体3基因的克隆及真核表达

采用RT-PCR方法从小鼠巨噬细胞中克隆小鼠Toll样受体3(TLR3)基因,基因测序表明获得了小鼠全长TLR3cDNA,构建了真核表达质粒p3XFLAG-CMV-7.1-TLR3.重组质粒转染293T细胞,Western blotting检测蛋白表达,表达蛋白质的相对分子量与预计相符.采用TLR3的阳性刺激物poly(I∶C)刺激重组质粒转染的293T细胞,双荧光素酶报告基因系统检测发现能激活下游转录因子NF-κB的转录活性,并能诱导TLR3下游细胞因子IL-6和TNF-α的表达.小鼠TLR3基因的克隆和表达,为研究TLR3介导的信号通路及其在抗病毒免疫中的作用打下基础.     

Chromosomal aberrations in bone marrow and peripheral blood cells from the same rats

Spontaneous cytogenetic aberrations were analyzed in bone-marrow cells and cultured peripheral lymphocytes from the same animals. No significant differences in the total number of cells with aberrations or total aberrations were detected between the bone-marrow cells and cultured lymphocytes. It was concluded that short-term culture does not contribute significantly to in vivo aberration yield within the experimental conditions used.     

Phenotypic comparison of multiple monocyte-related populations in murine peripheral blood and bone marrow.

BACKGROUND: Monocyte subsets are not well defined in murine peripheral blood (PB). Monocyte-related populations could also be located in bone marrow (BM), but studies correlating monocyte populations found in these two tissues are lacking. This study simultaneously analyzed PB and BM to phenotypically define multiple monocyte-related subsets in each location. METHODS: Murine PB and BM cells were simultaneously stained for monocyte-related populations, using five-color flow cytometry. Relevant subsets were defined on the basis of Ly-6C, CD11b, and wheat germ agglutinin phenotype in addition to light-scatter characteristics. These populations were extensively characterized for the expression of other myeloid and dendritic cell markers, adhesion molecules, chemokine receptors, and growth factor receptors. RESULTS: Six monocyte-related populations were defined, three each in BM and PB. No identical populations were found between the two tissues. Two BM populations and one PB population have heterogeneous expression of many markers, suggesting additional complexity among monocyte-related subsets. CONCLUSIONS: The murine monocytic series comprises multiple subsets, differing between PB and BM. This study defines and extensively phenotypes six of these populations, providing preliminary information about possible developmental relationships and migratory capacities of these cells.     

Exhaled breath profiling by electronic nose enabled discrimination of allergic rhinitis and extrinsic asthma

Aim: To assess whether an e-nose could discriminate between subjects affected by allergic rhinitis with and without concomitant extrinsic asthma, as well as from healthy controls, in terms of exhaled VOC-profile.

Methods: Fourteen patients with Extrinsic Asthma and Allergic Rhinitis (AAR), 14 patients with Allergic Rhinitis without asthma (AR) and 14 healthy controls (HC) participated in a cross-sectional study. Exhaled breath was collected by a standardized method and sampled by an e-nose (Cyranose 320). Raw data were reduced by Principal component analysis and analyzed by canonical discriminant analysis. Cross-validation accuracy (CVA) and Receiver Operating Characteristic(ROC)-curves were calculated. External validation in newly recruited patients (7 AAR, 7?AR and 7 HC) was tested using the previous training model.

Results: Breathprints of patients with AR clustered from those with AAR (CVA?=?85.7%), as well as HC (CVA?=?82.1%). Breathprints from AAR were also separated from those of HC (CVA?=?75.0%). External validation confirmed the above findings.

Conclusions: An e-nose can discriminate exhaled breath from subjects with allergic rhinitis with and without extrinsic asthma, which represent two different diseases with partly overlapping features. This supports the view of using breath profiling to diagnose asthma also in patients with allergic rhinitis.     

The effect of refrigeration of bone marrow and peripheral blood on cytogenetic analysis

Summary Bone marrow samples from patients with various hematologic disorders were stored at 4° C for up to 5 d before the establishment of a 24-h culture. We tested various factors, including storage time, colony stimulating factor, and methotrexate in an effort to improve metaphase and chromosome quality. Cytogenetic findings for various hematologic diseases were compared in a total of 201 cultures. Cold storage for up to 3 d did not seem to adversely affect the number of mitoses or the quality of chromosome banding when cells were cultured in a system that used both colony stimulating factor and methotrexate. In samples studies in parallel, clonal abnormalities were noted as frequently in cells stored in the cold as in those processed directly. This work was supported by contract DE-AC02-80EV10328 and by grant CA 16910 from the National Cancer Institute, Bethesda, MD.     

T-lymphocyte subpopulations in the peripheral blood and bone marrow of patients with aplastic anemia

A decrease in the absolute number of total lymphocytes, OKT3+ and OKT4+ lymphocytes, and a normal number of OKT8+ lymphocytes were found in the peripheral blood of patients with aplastic anemia. The OKT4:OKT8 ratio was decreased in patients due to a reduction in the percentage of OKT4+ cells and 3 out of 18 patients had a ratio less than 1. The values of the OKT4:OKT8 ratio were not associated either with the severity of the disease or with treatment with androgens. There was no correlation between the OKT4:OKT8 ratio and the number of transfusions received by patients. On the other hand, studies performed with bone marrow lymphocytes showed that the OKT4:OKT8 ratio for both patients and controls was lower than that of the peripheral blood. Since the ratio of OKT4:OKT8 cells in aplastic and control bone marrow was similar no direct pathogenic role can be assigned to the marrow for the imbalance detected in the peripheral blood.