Minor histocompatibility antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T-cell clones inhibit human hematopoietic progenitor cell growth by a mechanism that is dependent on direct cell-cell contact

HLA-identical bone marrow transplantation (BMT) may be complicated by graft-versus-host disease or graft rejection. Both complications are thought to be initiated by recognition of minor histocompatibility (mH) antigens by HLA-restricted mH-antigen-specific T lymphocytes. Using HLA- A2-restricted mH antigens HA-1-, -2-, and -4-, and HY-specific cytotoxic T lymphocyte (CTL) clones, we studied the recognition by these CTL clones of interleukin-2 (IL-2)-stimulated T cells (IL-2 blasts), BM mononuclear cells (BMMNCs), and hematopoietic progenitor cells (HPCs). We showed that, when IL-2 blasts from the BM donors who were investigated were recognized by the HA-1-, -2-, and -4-, and HY- specific CTL clones, their BMMNCs and HPCs were recognized as well by these CTL clones, resulting in antigen-specific growth inhibition of erythrocyte burst-forming units (BFU-E), colony-forming units- granulocyte (CFU-G), and CFU-macrophage (CFU-M). the HA-2-specific CTL clone, however, inhibited BFU-E and CFU-G growth from four donors to a lesser extent than from two other donors. We further investigated whether inhibitory cytokines released into the culture medium by the antigen-specific stimulated CTLs or by stimulated BMMNCs were responsible for suppression of HPC growth or whether this effect was caused by direct cell-cell contact between CTLs and HPCs. HPC growth inhibition was only observed after preincubation of BMMNCs and CTLs together for 4 hours before plating the cells in semisolid HPC culture medium. When no cell-cell contact was permitted before plating, neither antigen-stimulated CTL nor antigen-nonstimulated CTLs provoked HPC growth inhibition. Culturing BMMNCs in the presence of supernatants harvested after incubation of BMMNCs and CTL clones together for 4 or 72 hours did also not result in HPC growth inhibition. Both suppression of HPC growth and lysis of IL-2 blasts and BMMNCs in the 51Cr-release assay appeared to be dependent on direct cell-cell contact between target cells and CTLs and were not caused by the release of inhibitory cytokines into the culture medium by antigen-specific stimulated CTLs or by stimulated BMMNCs. Our results show that mH-antigen-specific CTLs can inhibit HPC growth by a direct cytolytic effect and may therefore be responsible for BM graft rejection after HLA-identical BMT.     

急性阑尾炎多层螺旋CT扫描征象与血清炎性标志物的相关性研究

目的探讨急性阑尾炎多层螺旋CT征象与血清炎性标志物的关系。方法收集2012年1月至2013年12月于同济大学附属杨浦医院经手术病理证实的具有完整临床及影像资料的急性阑尾炎患者66例,对所有患者的急性阑尾病变程度行CT分级,分析其与患者白细胞(WBC)计数、中性粒细胞百分比(NEUT%)及血清C反应蛋白(CRP)水平的相关性。结果急性阑尾炎CT分级与患者WBC计数与CRP水平呈正相关(P0.05),穿孔性阑尾炎患者NEUT%及CRP水平明显高于其他患者。CRP水平与阑尾直径、阑尾积液、回盲部变化、阑尾周围炎性条纹、小肠积液呈正相关(P0.05),WBC计数与回盲部变化及阑尾周围炎性条纹呈正相关(P0.05)。结论WBC计数及CRP水平与急性阑尾炎CT分级有关,CRP对急性阑尾炎的诊断及其严重程度的判断更有优势;CRP、NEUT%是穿孔性阑尾炎的重要预测因子,WBC可以更好地发现早期阑尾周围的炎性反应,综合分析CT表现与血清炎性标志物能更准确地诊断急性阑尾炎。     

肺炎支原体肺炎患儿肺泡灌洗液病菌量与临床特征相关性研究

目的研究肺炎支原体(MP)肺炎患儿肺泡灌洗液(BALF)中MP-DNA基因拷贝数和患儿病情严重程度的关系。方法选取河北省儿童医院2012年10月至2013年12月收治的82例MP肺炎患儿作为研究对象,行支气管镜下支气管肺泡灌洗(BAL),采用荧光实时定量聚合酶链反应(FQ-PCR)对肺泡灌洗液中MP-DNA定量检测,并根据检测结果的基因拷贝数分为低菌量组(MP-DNA的拷贝数103/mL的患儿),中等菌量组(MP-DNA的拷贝数为103~106/mL的患儿)和高菌量组(MP-DNA的拷贝数106/mL的患儿)。比较不同菌量组患儿的临床症状、实验室检查结果和影像学结果。结果高菌量组患儿总病程长,高热患儿和热程大于或等于7d的患儿数量均多于中、低菌量组,使用大环内酯类药物后退热时间也较其他两组更长,差异均具有统计学意义(P=0.027、P=0.025、P=0.029、P=0.003)。实验室检查中高菌量组C反应蛋白值升高明显,高于中低菌量组,差异有统计学意义(P=0.005)。影像学检查中高菌量组大片肺实变、肺不张者较、低菌量组多(P=0.002)。低菌量组未见双侧胸腔积液或大量胸腔积液患儿,中高菌量组此症状患儿较多,差异有统计学意义(P=0.033)。结论 MP肺炎患儿BALF病菌量和临床表现密切相关,高菌量组患儿病情更为严重。可能与患儿体内的肺炎支原体不易清除和存在较强的免疫反应有关,临床需延长抗菌药物治疗时间以及加强免疫治疗。     

肠球菌医院感染特征及新型抗菌药物耐药性研究

目的探讨肠球菌属细菌分布及耐药性特征,为指导临床合理用药及控制医院感染提供依据。方法对该院2011年1月至2013年12月年临床送检标本进行细菌分离培养、鉴定和药敏试验。结果共检出肠球菌属细菌140株,屎肠球菌71株(50.7%),粪肠球菌60株(42.9%),其他肠球菌9株(6.4%),其中尿液99株(70.7%);粪肠球菌对青霉素、氨苄西林、红霉素的耐药率为15.0%、12.5%和75.0%,屎肠球菌对青霉素、氨苄西林、左氧氟沙星、环丙沙星、红霉素耐药率大于80.0%,粪肠球菌和屎肠球菌对万古霉素(5.0%、4.2%)、利奈唑胺(8.4%、1.4%)极度敏感,喹奴普丁/达福普丁对粪肠球菌耐药率(100.0%)高于屎肠球菌(26.7%),达托霉素无耐药菌株。结论肠球菌属以泌尿系统感染为主,屎肠球菌检出率略大于粪肠球菌,屎肠球菌对大多数抗菌药物耐药率高于粪肠球菌,喹奴普丁/达福普丁仅对屎肠球菌有较高敏感性,万古霉素、利奈唑胺、达托霉素对肠球菌属细菌保持极高敏感性。     

16S rDNA测序技术在婴儿肠道微生态研究中的应用

目的探讨16SrDNA测序技术在新生儿、婴儿肠道微生态研究中的应用。方法于生后3天、1月、6月、1岁时收集2例健康婴儿粪便标本共8份,提取细菌总DNA,以Illumina Hiseq 2000为测序平台,采用新一代高通量16SrDNA宏基因组测序技术对V6可变区测序,并进行生物信息分析(物种分类和丰度分析;多样性分析)。结果 8份样品共产生原始测序数据为1 027.47 Mbp,Unique tags序列数量均值为58630,OTU数量63~209;优势菌门为Proteobacteria和Firmicutes;在科水平,1%的物种1个月之内2~4种,6月后达7~10种;1号婴儿一直以Enterobacteriaceae占优势,2号婴儿优势菌群包括Enterobacteriaceae、Lachnospiraceae、Streptococcaceae和Bacteroidaceae;4个时间点的npShannon和Simpson指数分别为1.17、1.29、2.16、2.51和0.43、0.40、0.26、0.14。结论 16SrDNA测序技术能满足新生儿、婴儿肠道微生态研究需求;新生儿、婴儿粪便中含丰富细菌基因组;细菌物种丰度及分类存在个体差异;从出生到1岁,婴儿肠道菌群结构趋向复杂和多样。     

Relations among Glycemic Control, Circulating Endothelial Cells, Nitric Oxide, and Flow Mediated Dilation in Patients with Type 2 Diabetes Mellitus

Circulating endothelial cells (CEC) have been identified as a surrogate marker of endothelial dysfunction. The aim of this study was to determine the association of glycemic control with CEC and endothelial function in patients with type 2 diabetes mellitus (DM).We studied 30 patients with type 2 DM and 20 age and sex matched healthy controls (HC). Number of circulating endothelial cells was measured by flow cytometry. Endothelial function was studied by measuring flow mediated vasodilation (FMD%) in the brachial artery and serum level of nitric oxide (NO).CEC count was significantly elevated in patients with DM, than HC (35.3±15.1 vs. 7.3±2.4, p<0.001) and in patients with HbA1c>7 than patients with HbA1c≤7 (47.4±5.5 vs. 19.5±5.7, p<0.001). FMD% and NO were lower in DM patients than HC (3.5±0.85 vs. 9.5±3.1, p<0.001 and 37.8±6.1 vs. 64.1±5.7, p<0.001 respectively). FMD% and NO were lower in patients with HbA1c>7 as compared to patients with HA1c≤7 (2.8±0.4 vs. 4.3±0.4, p<0.001 and 33.1±2.9 vs. 43.9±2.8, respectively, p<0.001). HbA1c correlated negatively with FMD% and NO levels and positively with CEC. CEC count correlated negatively with FMD% and NO. There was a significant positive correlation between CEC count and HBA1c (p<0.001 for all correlations).CEC is associated with markers of endothelial dysfunction and disease control in patients with type 2 DM. These findings suggest a potential role of CEC in the pathophysiology of cardiovascular disease in type 2 diabetic and raise the importance of tight glycemic control.     

Evidence for superantigen involvement in severe group a streptococcal tissue infections

Host-pathogen interactions were studied in tissue biopsy samples from patients with severe invasive group A streptococcus (GAS) infections. Skin, subcutaneous tissue, and fascia biopsy samples were divided into clinical grade 1 (no evidence of inflammation [n=7]) or clinical grade 2 (inflamed tissue--erythema and edema including cellulitis, fasciitis, and necrotizing fasciitis [n=24]). In situ imaging demonstrated significantly higher bacterial load in biopsy samples of higher clinical grade (P<.05), and the bacterial load correlated with the in vivo expression of the superantigen streptococcal pyrogenic exotoxin F (P<.02). Increased expression of the interleukin-1 cytokines and significantly higher expression of tumor necrosis factor-beta, interferon-gamma, and the homing receptors CC chemokine receptor 5, CD44, and cutaneous lymphocyte-associated antigen (P<.002-.05) were observed in biopsy samples of higher clinical grade. Thus, the cytokine profile at the local site of infection mimics that of a typical superantigen cytokine response. The findings of this study demonstrate a critical role for superantigens and Th1 cytokines in GAS tissue infections.     

A longitudinal immunologic evaluation of hemophiliac patients

Over an average span of one year, we performed a prospective clinical and immunologic evaluation of 30 patients with hemophilia. No patient developed life-threatening opportunistic infection or malignancy; however, the immunologic abnormalities and lymphadenopathy initially present in nine patients (lymphadenopathy group) persisted. In addition, five patients, representing 24% of the initial group without lymphadenopathy, developed generalized lymphadenopathy (converter group). One episode of idiopathic thrombocytopenia (ITP) and one episode of staphylococcal sepsis occurred in this "converter" group; one episode of ITP also occurred in the lymphadenopathy group. Sixteen patients remained asymptomatic. At the time of the follow-up evaluation, those differences in mononuclear cell (MNC) percentages and numbers noted initially among the three hemophiliac groups were no longer present. Natural killer cell function alone or in the presence of biologic response modifiers was not different among hemophiliac and control groups. Before developing lymphadenopathy, the converter group of patients had significantly better lymphocyte mitogenic function than did the other two groups of patients with hemophilia. However, lymphocyte mitogenic responses of all groups of patients with hemophilia significantly deteriorated over the course of the study. The abnormal mitogenic responses noted in these patients was explained in part by higher levels of spontaneous suppressor cell activity in mononuclear cell preparations from patients with hemophilia. We conclude that long-term immunologic studies of this patient population requires both quantitative and qualitative evaluations. Our data show that patients with hemophilia have progressive dysfunction of cell- mediated immunity.     

Modulation of expression of superantigens by human transferrin and lactoferrin: a novel mechanism in host-Streptococcus interactions

The role played by host-pathogen interactions in regulation of expression of streptococcal virulence factors in vivo is beginning to become clear. We have reported that the expression of 2 streptococcal virulence factors, the streptococcal pyrogenic exotoxin (Spe) A and the cysteine protease SpeB, was reciprocally modulated during infection with Streptococcus pyogenes. To identify host signals mediating this reciprocal regulation, we cocultured clonal M1T1 isolates with human peripheral blood mononuclear cells (PBMCs). In accordance with our in vivo findings, when bacteria were in direct contact with human PBMCs or were separated in transwells, expression of speA was induced, whereas expression of speB was down-regulated. This phenomenon was mediated by transferrin and lactoferrin and was influenced by the iron-saturation status of these proteins. Iron chelation from media induced expression of speA, but to a much lesser degree than did that with apotransferrin and lactoferrin, suggesting additional effects of these ferrins on modulation of expression of speA and speB. Thus, ferrins may play an important role in host-pathogen interactions in skin and mucosal tissues.