HLA-DR4 and salivary immunoglobulin A reactions to oral streptococci

The aim of this study was to describe and compare salivary immunoglobulin A (IgA) antibody reactions to extracts of strains of three oral streptococci in human leukocyte antigen (HLA)-DR4-positive and -DR4-negative subjects. Whole paraffin-stimulated saliva samples were collected from 27 apparently healthy subjects. Previous HLA typing showed that 20 subjects were DR4 positive and 7 were DR4 negative. HLA-DRB1*04 subtyping was performed among the DR4-positive subjects. Whole-cell antigen extracts from Streptococcus mutans (KPSK 2), Streptococcus sobrinus (OMZ 65) and Streptococcus parasanguis (Nt 62) were separated in SDS-PAGE. The antigens were immunoblotted with diluted saliva (Western blot), scanned and analyzed in a computer system. All immunoblot bands were recorded in DR4-positive and DR4-negative saliva pools, and bands with an optical density >or=0.1 were selected for analysis in individual salivas. The DR4-negative subjects in general had more immunoblot bands and more distinct bands than did the DR4-positive subjects. A higher concentration of total IgA in saliva was correlated with more bands, especially to antigens separated from S. mutans. When the number of bands was calculated per IgA unit, significant differences were observed between DR4-positive and DR4-negative salivas. This was particularly seen for S. mutans and S. parasanguis. As the number of bands was analyzed in relation to DR4 subgroups, DRB1*04, there was a lower salivary IgA activity to S. mutans in the DRB1*0401 and *0404. The variable level of correlation previously demonstrated for S. mutans colonisation and serologically defined DR4 positive subjects might be explained by the heterogeneity in this group, and the relation should be sought on a subgroup level.     

中国北方汉族人HLA-Ⅰ、Ⅱ类基因与HBV水平感染及预后相关性研究

目的探讨人类白细胞抗原Ⅰ、Ⅱ类基因与乙型肝炎病毒(HBV)感染及其预后的相关性。方法采用PCR-SSP方法分别对102名慢性持续性HBV感染者(实验1组)、76名急性HBV感染者(实验2组)及1383名中华骨髓库山东分库无血缘关系自愿骨髓捐献者(对照组)作HLA-Ⅰ、Ⅱ类基因分型检测,将结果作统计学处理。结果HLA-DR9、DR12在实验组(实验1组+2组)中的分布频率较对照组明显增高(P<0.01);HLA-DR8在实验1组的分布频率明显高于实验2组(P<0.01);HLA-DR13,DR15在实验1组中的频率明显低于实验2组(P<0.01,P<0.05)。结论HLA-DR9可能有助于HBV对宿主的感染;HLA-DR8,DR12可能有助于HBV对宿主的持续感染,对HBV感染的慢性化有促进作用;HLA-DR13,DR15可能有助于宿主对HBV的清除,对HBV感染的慢性化有抑制作用。     

目标血糖管理对脓毒症患者 CD69及 HLA －DR 表达的影响及其临床意义

目的：探讨早期目标血糖管理对脓毒症患者T淋巴细胞CD69、CD 14＋单核细胞（ HLA－DR）表达的影响及其临床意义。方法90例脓毒症患者按随机数字表法分为三组：血糖控制A组（4．4～6．1 mmol／L）30例、血糖控制B组（6．2～8．3 mmol／L）30例、血糖控制C 组（8．4～10．0 mmol／L）30例。入住ICU时就给予早期目标血糖管理,分别在入院时（0 d）及治疗后第1、3、7天采集静脉血,应用流式细胞仪检测T淋巴细胞CD69、CD14＋单核细胞HLA－DR的表达水平。相同时间点观察APACHEⅡ评分,并计算28 d病死率、MODS发生率及低血糖发生率。结果 A组第1、3、7天CD69和HLA－DR表达水平均较入院时明显升高,差异有统计学意义（ P均＜0.05）;B组第1、7天CD69表达水平较入院时明显升高,差异有统计学意义（P均＜0．05）,第3、7天HLA－DR表达水平与入院时比较差异有统计学意义（ P＜0．05）;C组CD69和HLA－DR表达呈先降低后升高的趋势,第3、7天CD69表达水平与入院时比较差异均有统计学意义（ P均＜0.05）,第1、7天HLA－DR表达水平与入院时比较差异有统计学意义（P＜0．05）。三组APACHEⅡ评分第1、3、7天较入院时均降低,差异有统计学意义（ P＜0．05）;第7天A组与B、C组比较显著降低,差异有统计学意义（P＜0．05）。三组28 d死亡率、MODS发生率及低血糖发生率均无统计学意义（ P均＞0．05）。结论目标血糖管理水平（4．4～6．1 mmol／L）能够促进CD69、HLA－DR的表达,有利于免疫稳态的恢复,可能对终止脓毒症患者免疫紊乱和免疫麻痹的恶性进程有利。     

HLA-DM recognizes the flexible conformation of major histocompatibility complex class II

DM facilitates formation of high affinity complexes of peptide-major histocompatibility complex (MHC) by release of class II MHC-associated invariant chain peptide (CLIP). This has been proposed to occur through discrimination of complex stability. By probing kinetic and conformational intermediates of the wild-type and mutant human histocompatibility leukocyte antigen (HLA)-DR1-peptide complexes, and examining their reactivities with DM, we propose that DM interacts with the flexible hydrophobic pocket 1 of DR1 and converts the molecule into a conformation that is highly peptide receptive. A more rigid conformation, generated upon filling of pocket 1, is less susceptible to DM effects. Thus, DM edits peptide-MHC by recognition of the flexibility rather than stability of the complex.     

Phenotype and TCR-gamma gene rearrangements in a Malaysian cohort of T-cell leukaemia/lymphoma cases

Abstract

T cells undergo a series of complex phenotypic changes before achieving maturation. Discrete stages of T-cell differentiation are simplified to four stages (pro-, pre-, cortical and mature-T cell) and used in the classification of T-cell leukaemia. HLA-DR has been reported to be expressed in immature T-cell acute lymphoblastic leukemia (ALL) and also confer a poorer treatment outcome. Simultaneously, the genotype goes through distinct pattern changes due to rearrangement of T-cell receptor (TCR) genes. TCR gene rearrangement is important in the diagnosis of clonality and used as markers to detect minimal residual disease in lymphoproliferative disorders. We identified a subset within Pro-T and Pre-T cell cases distinguished by the expression of HLA-DR. These subgroups appeared to be more immature as rearrangement of the TCR-gamma gene was either at germline or involved only the first constant region (C1) unlike a more rearranged pattern in the HLA-DR-subgroups. We also observed a higher incidence of mediastinal mass (67%) in the HLA-DR-subgroup in the Pre-T stage. These characteristics may be useful as markers to further refine staging of T-cell ALL and determine prognosis.     

Journal Watch

Abstract

Purpose: To correlate subclinical conjunctival inflammation and trabeculectomy results.

Methods: Prospective case series of 28 patients with primary open-angle glaucoma (28 eyes) under topical anti-glaucoma medication who underwent trabeculectomy. During surgery, a sample from the inferior bulbar conjunctiva was collected and the expression of HLA-DR together with the presence of inflammatory cells was correlated with trabeculectomy outcomes after 24 months. Surgical success was defined as intraocular pressure between 6 and 20?mmHg irrespective of the use of anti-glaucoma medication.

Results: Five patients missed follow-up visits and were removed from the study. Ten eyes (43.5%) were HLA-DR⁺, but no significant differences were observed between eyes with successful and failed surgeries (p?=?0.214). There was no significant association between the number of neutrophils and surgical outcomes (p?=?0.353).

Conclusions: The presence of inflammatory cells and expression of the inflammation marker HLA-DR in the conjunctiva did not correlate with the prognosis of trabeculectomy in this study.     

Limitations of an ocular surface inflammatory biomarker in impression cytology specimens

Context: A number of ocular conditions, such as dry eye, are associated with inflammation on the surface of the eye leading to irritation and ocular pain. Many drugs such as chemotherapeutics, beta blockers, angiotensin-converting enzymes and so forth also cause dry eye but currently there are no validated ocular surface biomarkers available.

Objective: We evaluated sample stability, assay sensitivity, reproducibility and overall performance of impression cytology (IC) utilizing the cellular surface biomarker human leukocyte antigen DR-1 (HLA-DR) as an ocular surface inflammatory biomarker by flow cytometry in a fit-for-purpose validation study. Additionally, subjects classified as normal or having various degrees of dry eye were evaluated to determine if HLA-DR could demonstrate a clear separation between normal and dry eye samples.

Results: The assay demonstrated high dynamic range detecting a broad range of fluorescent intensities in healthy donors. Additionally, inter, intra and stability assay results demonstrated strong concordance and low variability. Overall CV% for both assays were less than 25% for all measured parameters. However, high variability was observed for donor samples assayed beyond day 10 post IC sample collection (4.2–110.8 CV%).

Discussion: HLA-DR expression demonstrated a progressive increase in patients with mild to severe levels of dry eye disease providing sufficient evidence it is sensitive enough to monitor inflammatory effects of dry eye when coupled with additional biomarkers and/or methodologies such as cytokine analysis or ICAM-1. This biomarker can be used to monitor ocular surface disorders in patients and to evaluate potential treatment options during drug development. Although our results demonstrate this methodology is reproducible for routine evaluation, limitations around sample integrity exist.

Conclusion: The ocular cell surface inflammatory biomarker, HLA-DR coupled with impression cytology is a simple non-invasive robust, specific and reproducible assay that can be utilized to measure inflammatory infiltrates on the surface of the eye in IC samples less than 10-days old.     

检测组织相容性抗原与肝病的相关性

应用PAP免疫酶组织化学染色技术对各类乙肝患者的Mon、B、CD_3~+、CD_4~+、CD_8~+细胞表面HLA-DR、DQ抗原表达进行了检测,结果表明:各类乙肝患者DR~+Mon,DR~+B及DQ~+Mon、DQ~+B百分率均有不同程度的下降,以慢活肝、重症肝炎及肝硬化降低显著(P<0.001);DR~+CD_3、DQ~+CD_3百分率增高,以慢活肝、重症肝炎及肝硬化增高明显(P<0.001);DR~+CD_4、DQ~+CD_4百分率下降,DR~+CD_8、DQ~+CD_8百分率增高,以慢活肝、重症肝炎及肝硬化增高明显(P<0.001)。证明这些基因产物,在细胞之间相互作用的免疫应答及调节上起着关键性作用,与疾病的发生发展密切相关。     

A multistage-polyepitope vaccine protects against Mycobacterium tuberculosis infection in HLA-DR3 transgenic mice

Mycobacterium tuberculosis (Mtb) is responsible for almost 2 million deaths annually. BCG, currently the only TB vaccine, induces variable protection and does not protect against reactivation of latent TB. Thus, efficient vaccines to supplement BCG are required urgently. Since Mtb's proteome differs qualitatively and quantitatively during bacterial replication stages from that expressed during dormancy, improved TB vaccines should drive immune responses to Mtb antigens expressed during multiple stages of infection. Consequently, such “multistage” vaccines should be composed of (immunodominant) antigens expressed during different phases of Mtb infection. As a concept multistage vaccine, we constructed a polyepitope by fusing five HLA-DR3-restricted T-cell epitopes derived from different Mtb proteins either expressed highly by replicating bacteria (Ag85B, hsp65, 19 kDa lipoprotein), or abundantly expressed by dormant bacilli and recognized preferentially by TST⁺ individuals (hsp16, Rv1733c). PBMC of HLA-DR3⁺ but not HLA-DR3⁻ cured TB patients and TST⁺ individuals responded well to the multistage-polyepitope in vitro. The in vivo immunogenicity and protective efficacy of the multistage-polyepitope were analyzed using HLA-DR3 transgenic mice lacking endogenous murine class II as a model. Immunization with the multistage-polyepitope adjuvanted with CpG generated high IgG levels as well as polyfunctional CD4⁺ T-cells producing IFN-γ, TNF and IL-2, specific for these HLA-DR3-restricted epitopes. Importantly, multistage-polyepitope immunization reduced the number of bacilli in the lungs after Mtb challenge when administered as prophylactic vaccine. Given the extensive repertoire of potential Mtb antigens available for immune recognition, the data of our model demonstrate the potential of multistage-polyepitope vaccines to protect against TB.