A study of immunoglobulin G subclasses in patients with farmer's lung

A quantitative estimate of the IgG subclasses in patients with farmer's lung has revealed significantly higher IgG3 levels in patients with this condition compared to an age-matched group of control farmers similarly exposed to Micropolyspora faeni (P<0.01) and a group of normal blood bank donors (P < 0.001). The IgG1 and total IgG levels were significantly higher in both groups of farmers compared to blood bank controls (P<0.001). The possible significance of these observations is discussed in relation to susceptibility to the disease and its pathogenesis.     

Immunopathological studies in patients with farmer's lung

Immunohistological study of lung biopsies from four palients with farmer's lung revealed histological evidence of vasculilis along with deposits of immunoglobulins and C3 in and around the affected vessels in the biopsies from two palients. Both these patients were exposed to mouldy hay antigens shortly before biopsy. Both these patients also had considerable numbers of eosinophils in their lung lesions. Immune-complex mediated vasculitis is probably an early event in the pathogenesis of farmer's lung and might not be detectable in the absence of recent exposure to relevant antigens. The presence of large numbers of eosinophits in these lesions suggests a type I reaction acting as the trigger for a type 3 reaction.     

Distinct T cell subsets in adipose tissue are associated with obesity

Adipose tissue inflammation is a driving factor for the development of obesity-associated metabolic disturbances, and a role of adipose tissue T cells in initiating the pro-inflammatory signaling is emerging. However, data on human adipose tissue T cells in obesity are limited, reflected by the lack of phenotypic markers to define tissue-resident T cell subsets. In this study, we performed a deep characterization of T cells in blood and adipose tissue depots using multicolor flow cytometry and RNA sequencing. We identified distinct subsets of T cells associated with obesity expressing the activation markers, CD26 and CCR5, and obesity-specific genes that are potentially engaged in activating pro-inflammatory pathway, including ceramide signaling, autophagy, and IL-6 signaling. These findings increase our knowledge on the heterogeneity of T cells in adipose tissue and on subsets that may play a role in obesity-related pathogenesis.     

Immunohistochemical identification of exocrine and neuroendocrine subsets of large cell lung carcinomas

Formalin fixed, paraffin embedded sections of 52 cases of pulmonary large cell undifferentiated carcinoma (LCUC) as defined in the current WHO classification were studied immunohistochemically to assess features of exocrine and neuroendocrine (NE) differentiation. Monoclonal antibody 44-3A6 was applied to detect a membrane association protein related to exocrine differentiation. A panel of ten neuroendocrine markers including antibodies to synaptophysin, chromogranin A, serotonin, and seven neuropeptides was used to assess NE differentiation. The broad spectrum anticytokeratin antibody PKK1 was used to confirm the epithelial differentiation of these tumors. Exocrine differentiation was detected in 40/52 (77%) of surgically resected LCUC, despite the absence of recognizable glands by light microscopy. Eighteen of 52 (35%) LCUC exhibited NE differentiation; synaptophysin was the most frequently detected NE marker. Cytokeratin immunostaining with PKK1 was demonstrated in 41/52 (79%) cases. Subsets of LCUC were defined based on their expression of exocrine or NE phenotypic markers. Accordingly, 28/52 (54%) LCUC displayed an exocrine phenotype, 6/52 (12%) a NE phenotype, 12/52 (23%) had combined exocrine and NE phenotypes, and 6/52 (12%) exhibited neither phenotype. In this surgical series, there were no significant differences in stage at presentation for the four subsets. Interestingly, two year survival appeared decreased in patients with tumors displaying the "pure" NE phenotype.     

Serum immunoglobulin levels in farmer's lung disease

The serum concentrations of the five major classes of immunoglobulins were measured in 27 farmer's lung patients and compared with a group of normals and a group of patients with diffuse lung disease, but no precipitins to our antigen panel. The mean IgG and IgA levels in the sera of the farmer's lung patients were found to be significantly higher than that of the normal group. In addition, the IgG level of the farmer's lung group was also significantly higher than the group with negative precipitin tests. The remaining immunoglobulin classes, IgM, IgD, and IgE, were not remarkably different from normal in either diffuse lung disease group.     

Studies on the T-cell subset in lung tissue and BALF from patients with interstitial pneumonia]

The cellular components in bronchoalveolar lavage fluids (BALF) have been analyzed to obtain information on cellular kinetics in lung tissues of patients with diffuse pulmonary diseases. While various cells, alveolar macrophages, lymphocytes, and granulocytes appear in BALF, an increase in the percentage of lymphocytes has been noted in a variety of interstitial pneumonia. The T-cell subset of lymphocytes in BALF and biopsied lung tissues was, therefore, examined in patients with hypersensitivity pneumonitis (HP), sarcoidosis (Sar), idiopathic interstitial pneumonia (IIP) and rheumatoid arthritis with interstitial pneumonia (RA+IP) to compare the ratios of CD4/CD8 in BALF and lung tissues. The T-cell subset in BALF was analyzed by flow cytometry and the T-cell subset in lung tissues was detected with fresh frozen and thin-sliced specimens using an avidine-biotin complex (ABC) kit (Vecta Co. Ltd). The mean CD4/CD8 ratio in BALF was 0.36 in HP, 3.1 in Sar, 1.07 in IIP and 2.59 in RA+IP, while the mean CD4/CD8 ratio in lung tissues was 0.52 in HP, 2.59 in Sar, 1.11 in IIP and 2.25 in RA+IP, respectively. The ratios of CD4/CD8 in BALF and lung tissues from patients with these various interstitial pneumonia showed a positive correlation indicating that the changes of cellular components in BALF would reflect the changes in the lung tissues. Furthermore, CD4/CD8 ratios of lymphocytes infiltrating the alveolar portion and granuloma in lung tissues of HP were analyzed separately, because the CD4/CD8 ratios varied considerably from part to part of the lung tissues.(ABSTRACT TRUNCATED AT 250 WORDS)     

Novel phenotypes and migratory properties distinguish memory CD4 T cell subsets in lymphoid and lung tissue

Memory T cells are heterogeneous in expression of lymph node homing receptors, delineating "central-memory" (TCM, CD62Lhi/CCR7+) and "effector-memory" (TEM, CD62Llo/CCR7-) subsets that migrate to lymphoid and non-lymphoid tissues, respectively. It is not known how these subsets arise or how homing receptor expression and tissue origin determine their functional and migratory properties. Here, we investigated the role of CD62L expression in the generation, function, distribution and migration of heterogeneous memory CD4 T cells specific for influenza hemagglutinin (HA). We found that CD62Lhi and CD62Llo memory subsets are generated independent of CD62L expression by the activated precursor, and both subsets distribute into spleen and lung. Functionally, spleen- and lung-derived CD62L memory subsets produce effector cytokines at similar kinetics but differ strikingly in cell surface phenotype and migration: the CD62Llo memory subset expresses a classic memory phenotype (CD45RBlo/CD44hi/CD11a(hi)), while the CD62Lhi subset expresses an unconventional phenotype (CD45RBhi/CD44int/CD11a(int)), defining a new polyclonal memory subset. The CD62Lhi subset also trafficked more efficiently than CD62Llo cells into lymph nodes; however, only lung but not spleen CD62Llo memory T cells homed to lung. Our results reveal novel phenotypic heterogeneity of memory CD4 T cells co-segregating with CD62L expression and tissue-specific tropism of non-lymphoid memory CD4 T cells.     

小细胞肺癌90例免疫表型特征及其预后分析

目的探讨小细胞肺癌(small cell lung cancer,SCLC)神经内分泌标志物、CKpan及TTF-1与TNM分期及患者预后的关系。方法采用免疫组化Max Vision两步法检测90例SCLC组织中NSE、Cg A、Syn、CD56、CKpan及TTF-1的表达,分析患者的临床病理资料及TNM分期,电话随访计算患者的总生存期(overall survival,OS)。结果 90例SCLC中多数为老年男性,男女比为5∶1,中位年龄64岁,其中Ⅰ+Ⅱ期21例,Ⅲ期30例,Ⅳ期39例。SCLC中NSE、Cg A、Syn、CD56、CKpan及TTF-1的阳性率分别为83.3%、70%、65.5%、86%、92.2%和81.1%。Kaplan-Meier单因素分析显示TTF-1、NSE的表达与TNM分期及患者生存期相关(P<0.05);TTF-1阳性组中位生存期为8个月,与TTF-1阴性组(5.5个月)相比差异有统计学意义(P=0.000);NSE阳性组中位生存期为7个月,与NSE阴性组(11个月)相比差异有统计学意义(P=0.009);TNM分期中Ⅰ+Ⅱ、Ⅲ和Ⅳ的中位生存期分别为16个月、9个月和4个月,差异均有统计学意义(P=0.000);Cox多因素生存分析显示TTF-1表达和TNM分期是影响SCLC OS的独立预后因素。结论 SCLC多伴有神经内分泌分化,多数表达CKpan和TTF-1;其中TTF-1表达与SCLC的预后呈负相关;NSE表达与预后呈正相关;TTF-1表达和TNM分期是影响SCLC OS的独立预后因素。     

Double phenotyping of immunoregulatory T cell subsets in patients with allergic asthma

In order to determine whether the dissection of helper/inducer (CD4 +) and suppressor/cytotoxic (CD8 +) lymphocyte subsets with Leu 8 reagent would reveal any differences between allergic asthma patients and non-atopic controls, we compared in both groups the 'true helper' T cell subset (Leu 8 - CD4 +), responsible for the major helper effect, and one of the suppressor T cell subpopulations (Leu 8 - CD8 +). Peripheral blood mononuclear cells from sixty-nine individuals, including nineteen extrinsic asthmatics, fifteen intrinsic asthmatics, seventeen patients with chronic obstructive lung disease and eighteen healthy controls, were comparatively analysed. Although total CD4 + cells and total CD8 + cells were similar for all groups, we found in the extrinsic asthma patients group a significant increase in the number of 'true helper' T cell sublineage (Leu 8 - CD4 +) and of suppressor cells expressing Leu 8 - CD8 + phenotype. Such imbalances may be implicated in the pathogenesis of atopic asthma.     

The distribution and function of human memory T cell subsets in lung cancer

The distribution and function of T lymphocytes in human lung cancer remain limited. In this study, we investigated the properties of human T cell subsets in the blood of non-small cell lung cancer (NSCLC) patients. We found a relatively normal level of CD4+ subsets in the blood of NSCLC patients, but CD8+ effector T cells increased and CD8+ effector memory cells declined compared to the healthy donors. To further analyze their properties, we stimulated the peripheral blood mononuclear cells (PBMCs) of NSCLC patients by mitogens to examine cytokine production. Our data suggest that both CD4+ and CD8+ naïve cells in NSCLC patients significantly reduced IFN-γ and TNF-α production. Additionally, fewer CD8+ effector cells produced IFN-γ and TNF-α in NSCLC patients than in healthy subjects. Moreover, similar results were observed for CD4+ or CD8+ memory cells in NSCLC patients for the production of IFN-γ, TNF-α, and IL-17. Therefore, our results strongly suggest that the function of CD4+ and CD8+ T lymphocytes in NSCLC patients is compromised or dysregulated. The development of vaccines and antitumor immunotherapy may be essential for the treatment of lung cancer patients.     

帕金森病患者NK细胞亚群及T淋巴细胞亚群变化的临床意义

目的:探讨帕金森病(PD)不同年龄、不同病期、伴随症状患者外周血NK细胞亚群及T淋巴细胞亚群的变化及其临床意义。方法:应用先进的流式细胞仪(FCM)直接免疫荧光染色法检测47例PD患者外周血NK细胞亚群及T淋巴细胞亚群,并与健康人组进行对照与相关分析。结果:PD组CD3+、CD4+、CD8+、CD4+/CD8+水平较对照组均明显降低(P<0.05),而CD16+56水平则较对照组明显增高(P<0.05)。高龄、病情重及伴痴呆和抑郁的PD患者CD3+、CD4+、CD8+、CD4+/CD8+水平降低更显著(P<0.05)。结论:PD发病过程中存在T细胞免疫功能低下及NK细胞免疫平衡失调,高龄、病情重及伴痴呆和抑郁的患者NK、T细胞免疫功能异常降低更明显,此为PD病理生理基础赋予新的内涵,亦为PD的免疫干预性治疗提供新的途径。     

T cell subsets in patients with acute and chronic HBV infection,primary biliary cirrhosis and alcohol induced liver disease

The proportions of inducer and cytotoxic/suppressor T cells and their concentrations in peripheral blood have been determined in patients with acute and chronic type B hepatitis, hepatitis B virus (HBV) carriers with normal hepatic histology, patients with alcohol-induced liver disease (ALD), primary biliary cirrhosis (PBC) and chronic extrahepatic cholestasis. During acute type B hepatitis the inducer/suppressor ratio was decreased due to an increase in suppressor cell concentrations. When this ratio returned to normal the HBs antigen was cleared and HBs antibody was detectable. Similar abnormalities were found in patients with HBs + ve chronic hepatitis. In HBs antigen-positive patients with normal histology, normal T cell subsets were found.In some patients with primary biliary cirrhosis the ratio of inducer to suppressor cells was low due to a reduction in the concentration of inducer cells and in others high due to a reduction in suppressor cells. Administration of cyclosporin A to the latter group produced an increase in the concentration of suppressor cells and there was an improvement in liver biochemistry.In alcohol-induced hepatitis and cirrhosis the ratio of inducer/suppressor cells was normal.Whether these imbalances of the regulatory cells of the immune system in patients with chronic HBV-induced hepatitis and PBC are of primary or secondary importance is uncertain. The relationship of the depressed ratio to persistence of the hepatitis B virus is worthy of further study.     

HIV/AIDS患者外周血T淋巴细胞亚群的研究

目的 通过对HIV/AIDS患者外周血T淋巴细胞亚群进行分析,探讨初始、记忆和效应T细胞各亚群的变化情况及其与疾病进展的关系.方法 应用流式细胞仪检测15例正常人,79例HIV/AIDS患者CD4<200组17例、200≤CD4≤500组45例和CD4>500组17例.外周血淋巴细胞中T细胞各亚群绝对数及百分比.结果随着疾病进展,CD4+ 初始细胞(Naive)计数和比例均逐渐减少(P<0.001);CD4+中枢性记忆T细胞(Tcm)计数逐渐降低(P<0.001),但百分比逐渐升高(P=0.002);CD4+效应记忆性T细胞(TEMA)百分比上升(P<0.001);CD8+Naive细胞计数及百分比均逐渐下降(P<0.05);CD8+ TCM、TEM和终末分化的效应性记忆T细胞(TEMRA)的计数及百分比,各组间差异均无统计学意义(P>0.05).结论 HIV感染者外周血T淋巴细胞亚群发生显著变化,幼稚型T淋巴细胞数目逐渐减少,功能型T淋巴细胞数目增加.本研究有助于对HIV致病机制的研究及疾病进展的监测.     

强直性脊柱炎患者外周血T细胞亚群及相关免疫分子的变化

为探究强直性脊柱炎(ankylosing spondylitis,AS)的发病机制和免疫细胞在病情发展中的功能和重要性,我们设计本实验以比较强直性脊柱炎患者与正常人外周血中免疫细胞格局的变化。实验以42例AS患者和42例健康者为研究对象,检测外周血T细胞以及相应的细胞因子和转录因子。实验采用流式细胞术检测T细胞亚群的比例、通过ELISA方法检测细胞因子并使用Q-PCR检测转录因子的表达。实验发现,与健康人相比,AS患者外周血T细胞中CD4~+细胞占优势,并且effect T细胞与na?ve T细胞的比值升高;对CD4~+T细胞亚群进一步的检测结果显示Th1与Tfh的比例显著增高,Treg细胞的比例较正常人稍高。在对细胞因子的检测中,我们发现IFN-γ、TGF-β显著下降,但IL-21有上升趋势;对转录因子的检测显示,Bcl-6/Foxp3、RORγt/Foxp3和T-bet/Foxp3值均升高。上述结果提示强直性脊柱炎患者外周血中T细胞亚群的格局发生了变化,其相关免疫分子也呈现免疫活化状态。     

Regulatory T cell subsets in peripheral blood of celiac disease patients and TLR2 expression: correlation with oxidative stress

The study was carried out to study expression of Toll like receptors 2 (TLR2), natural/inducible Treg and Interferon‐γ alongside oxidative stress and understand their significance in pediatric samples. Influence of oxidative stress on Celiac Disease was analysed by evaluating lipid peroxidation, reduced glutathione, Glutathione peroxidase, etc. A comparison was performed among CD patients, CD patients on gluten free diet (GFD), and healthy controls. Peripheral nTregs exhibited a similar pattern of reduced numbers in CD and GFD cases when compared to healthy controls. On the other hand, inducible Tregs were much lower in GFD patients as compared to CD patients. Expression of TLR2 on iTregs was elevated in CD and GFD, however, expression on nTregs was unaltered in all the three groups. The inflammatory cytokine IFN‐γ positive Treg cells were found to be elevated in CD as compared to control group. Oxidative stress was elevated in CD as compared to healthy controls while that in GFD samples was lower in comparison to CD. The levels of LPO, activities of enzymes SOD and Catalase were higher in CD and GFD samples when compared to controls. However, enzyme Glutathione peroxidase and reduced glutathione levels declined in both CD and GFD groups as compared to controls. This report highlights the effect of elevated oxidative stress in CD on reduced traffic of iTregs toward periphery. A strong correlation was observed between the cytokine IFN‐γ and TLR2 expression in movement of iTregs in CD patients.     

Correlation of exposure to various respiratory pathogens with farmer's lung disease.

Complement-fixing antibodies (CFA) to a panel of microorganisms commonly associated with respiratory disease were measured in a number of agricultural populations. The panel included Mycoplasma pneumoniae, influenza viruses A and B, parainfluenza virus types 1, 2, and 3, adenovirus, and respiratory syncytial virus. The agricultural populations were grouped according to a clinical history of farmer's lung disease (FLD) and the presence of antibodies to the thermophilic actinomycetes (TA). Farmers with precipitating antibody activity to one or more of the TA (groups I and II) demonstrated a greater frequency of CFA to M. pneumoniae and parainfluenza virus types 1, 2, and 3 than those groups without antibody to the TA, but the level of CFA was not higher in the positive subjects. Immunoglobulin levels were also elevated in groups I and II when compared to the control groups. Unlike IgG and IgM, IgA was elevated only in the farmers who had a clinical history of FLD (group I) but not in farmers without a clinical history. The results suggest that farmers who develop FLD are exposed to a wider variety of pathogens than are other farmers, but do not respond in an accelerated manner.     

Immunoregulatory T cell subsets and T cell activation in rheumatoid arthritis

Conclusion Activation of T cell may be accomplished following the interaction of the TCR-CD3 complex with antigen and MHC products. In vitro this may be replaced by antibodies to the TcR or CD3 complex which mimic ligand binding. So called alternative pathways may also trigger activation. Activational state may be measured by lymphokine production, proliferative capacity or by expression of activation antigens. By these criteria a proportion of T cells isolated from rheumatoid joints appear to have undergone an in vivo activation. Phenotypic analysis of the synovial T cells has also established that there is an unusual distribution of T and T cells and T cell subsets in many rheumatoid patients. As T cells play a central role in immunoregulation, further exploitation of these observations using T cell clones and molecular techniques will extend our understanding of the disease process. In particular, further knowledge is required on the possible role of T cells in RA, the clonality of the T cells, the possible use of alternative activation pathways, and ultimately, the specificity of these T cells.