HLA-DRB1~* 1501与我国北方汉族儿童结核病的易感性研究

为了研究中国北方儿童结核病与HLAⅠ类基因的关联,我们采用PCR-SSO方法检测了97例北方汉族结核患儿和91例正常对照的HLA-DRBl,DQAl,DQBl等位基因.发现中国北方汉族儿童结核病与HLA-DRBl·1501有显著关联.进一步比较DR分子结构发现β链第86位氨基酸对结核病的易感性可能有重要意义.     

IL-10 down-regulates costimulatory molecules on Mycobacterium tuberculosis-pulsed macrophages and impairs the lytic activity of CD4 and CD8 CTL in tuberculosis patients

Activation of T cells requires both TCR-specific ligation and costimulation through accessory molecules during T cell priming. IFNgamma is a key cytokine responsible for macrophage activation during Mycobacterium tuberculosis (Mtb) infection while IL-10 is associated with suppression of cell mediated immunity in intracellular infection. In this paper we evaluated the role of IFNgamma and IL-10 on the function of cytotoxic T cells (CTL) and on the modulation of costimulatory molecules in healthy controls and patients with active tuberculosis (TB). gamma-irradiated-Mtb (i-Mtb) induced IL-10 production from CD14(+) cells from TB patients. Moreover, CD3(+) T cells of patients with advanced disease also produced IL-10 after i-Mtb stimulation. In healthy donors, IL-10 decreased the lytic activity of CD4(+) and CD8(+) T cells whereas it increased gammadelta-mediated cytotoxicity. Furthermore, we found that the presence of IL-10 induced a loss of the alternative processing pathways of antigen presentation along with a down-regulation of the expression of costimulatory molecule expression on monocytes and macrophages from healthy individuals. Conversely, neutralization of endogenous IL-10 or addition of IFNgamma to either effector or target cells from TB patients induced a strong lytic activity mediated by CD8(+) CTL together with an up-regulation of CD54 and CD86 expression on target cells. Moreover, we observed that macrophages from TB patients could use alternative pathways for i-Mtb presentation. Taken together, our results demonstrate that the presence of IL-10 during Mtb infection might contribute to mycobacteria persistence inside host macrophages through a mechanism that involved inhibition of MHC-restricted cytotoxicity against infected macrophages.     

Th1型细胞因子基因对结核分枝杆菌基因疫苗诱导BALB/c小鼠产生抗CFP10抗体水平的影响

目的：研究分别表达含IL—12和IL-18基因的质粒，对结核分枝杆菌(Mycobacterium tuberculosis，MTB)H37R1株CFP1O基因疫苗诱导免疫应答的影响。方法：从正常人外周血单个核细胞(PMBCs)中提取RNA，用RT—PCR扩增IL-18 cDNA，并克隆人载体pGEM—Teasy中。测序证实后，亚克隆至真核表达载体pcDNA3．1的BamH Ⅰ和EcoR Ⅰ酶切位点。将分别表达小鼠IL—12和人IL—18基因的真核表达质粒pcmlL12和pclL18，与MTB CFP10基因疫苗联合肌注免疫BALB／c小鼠，共免疫3次，每次间隔2wk。每次免疫后2wk采血、分离血清，用ELISA检测小鼠血清抗CFP10抗体的滴度。结果：用RT—PCR成功地从人PMBC的RNA中扩增出IL—18 cDNA，测序结果正确，用BamH Ⅰ和EcoR Ⅰ酶切鉴定证实，目的基因已插入载体pcDNA3．1中，阳性克隆命名为pcIL18。pcCFP10组第1次免疫后，血清抗CFP10抗体的平均滴度为1：600，末次免疫后的滴度为1：4000。pcIL18 pcCFP10组联合免疫后，血清抗CFP10抗体的滴度高于pcCFP10组，最终达1：8000。而pcmIL12 pcCFP10组联合免疫后滴度仅为1：200。结论：pcIL18与CFP10基因疫苗联合免疫，可增强CFP10抗原的特异性体液免疫应答；pcmIL12则可使CFP10基因疫苗产生的抗体水平降低。pcIL18 pcCFP10基因联合免疫是否具有增强CFP10抗原特异性细胞免疫的作用有待进一步研究。     

Opsonizing antibodies (IgG1) up-regulate monocyte proinflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and IL-6 but not anti-inflammatory cytokine IL-10 in mycobacterial antigen-stimulated monocytes-implications for pathogenesis

Cachexia is one of the prominent features of advanced tuberculosis (TB) seen in association with increased expression of the monokine TNF-alpha. Several mycobacterial proteins, including PPD, stimulate TNF-alpha secretion from monocytes. Host factors that may play a role in cytokine expression from monocytes remain largely unknown. One such factor is the opsonizing antibodies. Monocytes have high-affinity receptors (FcgammaI and FcgammaIII) for IgG1 and IgG3 antibodies that mediate antigen uptake. We have reported selective up-regulation of IgG1 (which bind to Fcgamma receptors) in advanced TB and have recently shown the ability of PPD-specific IgG1 antibodies to augment TNF-alpha expression in PPD-stimulated monocytes. These observations have now been extended to other cytokines with semipurified fractions from secreted antigens of Mycobacterium tuberculosis (containing 30 kD and 58 kD) that were devoid of lipids, glycolipids and carbohydrates. In the presence of heat-inactivated TB plasma containing known amounts of antigen-specific IgG1 antibodies, these fractions induced significantly increased TNF-alpha, IL-6 and IL-10 secretion. Absorption of IgG1 with Protein 'A' removed the augmenting activity for TNF-alpha and IL-6 secretion from the TB plasma samples. In the case of IL-10, removal of IgG1 resulted in increased rather than decreased IL-10 secretion. These results suggest a possible pathogenic role for antibodies in TB by enhancing proinflammatory and blocking down-regulatory cytokines such as IL-10 cytokines during the chronic phase of TB.     

Killing of Mycobacterium tuberculosis within human monocytes: activation by cytokines and calcitriol.

Human monocytes were isolated and their ability to harbour growth of virulent tubercle bacilli was assessed, in the presence or absence of various immunomodulators. Calcitriol (1,25(OH2), vitamin D3) alone, at doses of 10(-7)-10(-9) M endowed human monocytes with a significant ability to restrict intracellular growth of the tubercle bacilli. Crude immune lymphokines as well as recombinant interferon-gamma (IFN-gamma) endowed monocytes with no tuberculostatic activity. Similarly, other recombinant cytokines tested, notably colony-stimulating factor-1 (CSF-1), interleukin-1 (IL-1), interleukin-3 (IL-3) and interleukin-6 (IL-6) all failed to stimulate anti-tuberculous properties, and even increased growth of the tubercle bacilli in monocytes, in the case of CSF-1. Conversely, incubation of crude lymphokines in combination with calcitriol led to total stasis of the growth of M. tuberculosis. Experiments with recombinant cytokines and immunologically active vitamins showed that a combination of IFN-gamma tumour necrosis factor-alpha and calcitriol induced a significant amount of intramonocyte killing of M. tuberculosis. Addition of this cocktail of factors to already infected monocytes led to substantial killing of tubercle bacilli. These sets of experiments establish clearly that combinations of recombinant cytokines and vitamins may induce substantial intramonocyte killing of M. tuberculosis. The mechanism involved in this killing activity was not clarified.     

Malignant lymphoma of bronchus-associated lymphoid tissue (BALT) coexistent with pulmonary tuberculosis

A case in which malignant lymphoma occurred in association with a tuberculosis focus in a 70-year-old man is reported. Surrounding the epithelioid cell granulomas with caseous necrosis was a dense and diffuse monotonous infiltration of atypical lymphoid cells. Acid-fast bacilli were found in the granulomas and pulmonary tuberculosis was diagnosed. The infiltrating atypical lymphoid cells occasionally invaded the respiratory epithelium producing lymphoepithelial lesions. Immunohistochemically, the lymphoid cells were positive for CD20, and clonal rearrangement of the immunoglobulin heavy chain gene was demonstrated by polymerase chain reaction (PCR). We diagnosed the lesion as a pulmonary malignant lymphoma of bronchus-associated lymphoid tissue (BALT) occurring in the background of tuberculosis. This is the first reported case of pulmonary BALT lymphoma coexistent with pulmonary tuberculosis.     

青海省2011—2020年60岁以上老年肺结核患者就诊延迟现状及影响因素分析

目的 分析青海省2011—2020年老年肺结核患者就诊延迟现状及影响因素,为今后科学有效防治老年结核病提供科学依据。方法 分析描述青海省2011—2020年老年肺结核患者就诊延迟现状及变化趋势,采用单因素卡方检验、多因素logistic回归对老年肺结核患者就诊延迟影响因素进行分析。结果 青海省2011—2020年老年肺结核患者10004例,就诊延迟6146例,就诊天数中位数为22（8,56）,就诊延迟率为61.44%;2011—2020年,整体呈下降趋势（x2趋势=427.926,P<0.001）,不同性别（男性x2趋势=243.848,P<0.001;女性x2趋势=185.402,P<0.001）和年龄组（≤<70岁x2趋势=281.925,P<0.001;70~80岁x2趋势=123.219,P<0.001;>80岁x2趋势=20.175,P<0.001）患者就诊延迟率也均呈下降趋势;多因素Logistic回归分析结果显示,与汉族、本地户籍、因症就诊、Ⅰ型肺结核患者相比,其他少数民族（OR=1.308, 95% CI: 1.177~1.454）、因症推荐（OR=1.392, 95% CI: 1.108~1.749）、Ⅱ型（OR=3.314, 95% CI: 2.071~5.302）、Ⅲ型（OR=3.149, 95% CI: 2.104~4.713）、Ⅳ型（OR=2.695, 95% CI: 1.741~4.171）患者发生就诊延迟风险较高,流动人口（OR=0.707, 95% CI: 0.631~0.792）、转诊（OR=0.658, 95% CI: 0.597~0.726）、追踪（OR=0.873, 95% CI: 0.778~0.980）患者发生就诊延迟风险较低,差异均有统计学意义（P<0.05）。结论 青海省2011—2020年老年肺结核患者就诊延迟率较高,民族、户籍、患者来源和诊断分型是就诊延迟的影响因素。     

毕节市敏感肺结核患者家庭灾难性支出情况分析

目的 了解毕节市敏感肺结核患者家庭疾病灾难性支出（CTC）及影响因素,为降低患者家庭负担提供政策建议。方法 以2020年5—6月期间毕节市登记并在调查时已成功治疗的敏感患者为调查对象,通过概率比例抽样对其进行问卷访谈。灾难性支出采用率描述,用χ2检验和二分类logistic回归分析其原因。结果 毕节市309例肺结核患者家庭总自付费用 M （ P 25, P 75）为5 169.31（2 098.52,14 778.49）元;直接医疗费用、直接非医疗费用和间接费用 M （ P ? P ?分别为1 688.15（987.31,3 337.50）、720.00（200.00,1 425.00）和1 200.00（0.00,8 390.00）,差异具有统计学意义（χ2=63.545, P ＜0.001）。309例患者家庭CTC发生率为52.43%;其中,家庭年收入较低（ OR =25.740,95% CI :9.676～68.473）、住院（ OR =3.515,95% CI :1.782～6.933）、确诊延迟（ OR =2.492,95% CI :1.297～4.789）是导致其发生CTC的危险因素（ P 均＜0.05）。结论 毕节市肺结核患者经济负担以间接费用为主,CTC发生率较为普遍。其中,家庭年收入低、住院和确诊延迟患者是其重要危险因素,可针对性地加强患者健康教育、落实医疗机构诊疗规范以及通过政府和社会给予患者补偿,共同降低其疾病负担减少CTC情况。     

肺结核专科护理质量评价指标体系的构建

目的：构建肺结核专科护理质量评价指标体系,为肺结核临床护理质量管理提供参考依据。方法：以Donabedian的“结构–过程–结果 ”三维质量评价模型为理论基础,通过文献检索、德尔菲专家函询法、层次分析法确立肺结核专科护理质量评价指标体系。结果：共有45名专家完成两轮函询,问卷有效回收率均为100%;专家权威系数（Cr）分别为0.822和0.859;两轮函询肯德尔协调系数（Kendall’s W）分别为0.140～0.379和0.107～0.665,P均<0.01。根据条目筛选标准、专家意见及院内外专家小组会议讨论结果,确立终版评价指标体系含一级指标3项,二级指标18项,三级指标73项。结论：肺结核专科护理质量评价指标体系结构合理,构建过程科学、可靠,可为肺结核专科护理质量管理及专业学科发展提供理论参考。