Fluorescence In Situ Hybridization to Visualize Genetic Abnormalities in Interphase Cells of Acinar Cell Carcinoma, Ductal Adenocarcinoma, and Islet Cell Carcinoma of the Pancreas

OBJECTIVE: To use fluorescence in situ hybridization (FISH) to visualize genetic abnormalities in interphase cell nuclei (interphase FISH) of acinar cell carcinoma, ductal adenocarcinoma, and islet cell carcinoma of the pancreas.PATIENTS AND METHODS: Between April 4, 2007, and December 4, 2008, interphase FISH was used to study paraffin-embedded preparations of tissue obtained from 18 patients listed in the Mayo Clinic Biospecimen Resource for Pancreas Research with a confirmed diagnosis of acinar cell carcinoma, ductal adenocarcinoma, islet cell carcinoma, or pancreas without evidence of neoplasia. FISH probes were used for chromosome loci of APC (see glossary at end of article for expansion of all gene symbols), BRCA2, CTNNB1, EGFR, ERBB2, CDKN2A, TP53, TYMP, and TYMS. These FISH probes were used with control probes to distinguish among various kinds of chromosome abnormalities of number and structure.RESULTS: FISH abnormalities were observed in 12 (80%) of 15 patients with pancreatic cancer: 5 of 5 patients with acinar cell carcinoma, 5 of 5 patients with ductal adenocarcinoma, and 2 (40%) of 5 patients with islet cell carcinoma. All 3 specimens of pancreatic tissue without neoplasia had normal FISH results. Gains of CTNNB1 due to trisomy 3 occurred in each tumor with acinar cell carcinoma but in none of the other tumors in this study. FISH abnormalities of all other cancer genes studied were observed in all forms of pancreatic tumors in this investigation.CONCLUSION: FISH abnormalities of CTNNB1 due to trisomy 3 were observed only in acinar cell carcinoma. FISH abnormalities of genes implicated in familial cancer, tumor progression, and the 5-fluorouracil pathway were common but were not associated with specific types of pancreatic cancer.5FU = 5-fluorouracil; FISH = fluorescence in situ hybridization; ND-FISH = interphase FISH to detect abnormalities of chromosome number and structure; SSC = standard saline citratePancreatic cancer afflicts more than 200,000 new patients worldwide each year, including more than 37,600 in the United States.^1-4 Surgery seldom cures pancreatic cancer and effective chemotherapies are largely unknown.⁵ Survival varies among patients with pancreatic cancer but is often measured in months.^6,7 Thus, novel genetic research of pancreatic cancer is urgently needed to help establish accurate diagnoses and to develop effective treatments for this important public health problem.This investigation used fluorescence in situ hybridization (FISH) to visualize chromosomal loci in interphase nuclei (interphase FISH) of acinar cell carcinoma, ductal adenocarcinoma, and islet cell carcinoma of the pancreas. Acinar cell carcinoma and ductal adenocarcinoma, which are nonendocrine tumors, and islet cell carcinomas, which are neuroendocrine tumors, reportedly occur in less than 1%, 95%, and 5%, respectively, of patients with pancreatic cancer.^3,8,9 In 2 retrospective studies of large series of patients, median survival for unresected ductal adenocarcinoma was 3 months⁸; for unresected acinar cell carcinoma, 25 months⁸; for nonfunctional islet cell carcinoma, 26 months⁹; and for functional islet cell carcinoma, 54 months.⁹Although some genetic studies of pancreatic cancer have been published, most focus on ductal adenocarcinoma. An assortment of genetic procedures have been attempted to investigate pancreatic cancer, including family history studies,¹ conventional cytogenetic techniques,¹⁰ interphase FISH,^11-13 comparative genomic hybridization,¹⁴ RNA expression analyses,^15,16 and evaluation of sequence variations.¹⁷ These studies implicate a variety of point mutations and chromosomal anomalies in most, if not all, patients with pancreatic cancer. Nevertheless, the biology of various forms of pancreatic cancer remains poorly understood, and this problem precludes efforts to establish novel genetic treatments for these disorders.This investigation used interphase FISH to visualize genetic abnormalities in individual normal and neoplastic cells in acinar cell carcinoma, ductal adenocarcinoma, and islet cell carcinoma of the pancreas.¹⁸ This research project was specifically designed to detect genetic abnormalities that have been shown via other genetic technologies to be associated with acinar cell carcinoma of the pancreas, familial pancreatic cancer, tumor progression in pancreatic cancer, or genes linked with the molecular pathway of 5-fluorouracil (5FU) chemotherapy.     

Ultrasound Image Classification of Ductal Carcinoma In Situ (DCIS) of the Breast: Analysis of 705 DCIS Lesions

The Japan Association of Breast and Thyroid Sonology (JABTS) proposed, in 2003, a conceptual classification system for non-mass abnormalities to be applied in addition to the conventional concept of masses, to facilitate detecting ductal carcinoma in situ (DCIS) lesions. The aim of this study was to confirm the utility of this system and to clarify the distribution of these findings in DCIS lesions. Data on 705 surgically treated DCIS lesions from 16 institutions in Japan were retrospectively reviewed. All 705 DCIS lesions could be classified according to the JABTS classification system. The most frequent findings were hypo-echoic areas in the mammary gland (48.6%), followed by solid masses (28.0%) and duct abnormalities (10.2%) or mixed masses (8.1%). Distortion (1.3%), clustered microcysts (1.4%) and echogenic foci without a hypo-echoic area (2.5%) were uncommon. These results suggest that the concept of non-mass abnormalities is useful in detecting DCIS lesions.     

致敏的脐血源树突状细胞诱导CTL特异杀伤白血病细胞的实验研究

探讨脐血单个核细胞（MNC）诱导的树突状细胞（DC）通过负载冻融的HL-60、K562细胞抗原体外诱导产生细胞毒性T淋巴细胞（CTL）对HL-60、K562的杀伤作用。取脐血12份,分离MNC。在MNC中加入细胞因子GM-CSF（granulocyte monocyte colony-stimulating factor）、IL-3（interleukin 3）、SCF（stemcell factor）和EPO培养4周。使用CD83、CD1a、CD11C和CDw123单克隆抗体、流式细胞仪测定培养前后脐血DC抗原变化及扩增情况。DC通过负载HL-60、K-562白血病细胞抗原致敏T淋巴细胞产生CTL^3H-TdR掺入试验测定DC免疫刺激活性,MTT法观察CTL对HL-60、K562细胞的特异性杀伤活性。结果表明：新鲜脐血CD1a^＋、CD11c^＋、CD83^＋、CDw123^＋细胞数分别为0．27×10^5／ml、5．87×10^5／ml、1．94×10^5／ml、2．73×10^5／ml。加入上述细胞因子培养的脐血MNC分化为CD1a^＋、CD11C^＋、CD83^＋、CDw123^＋DC,经培养2—4周,DC数明显增多,分别达11．02×10^5／ml、28．24×10^5／ml、10．57×10^5／ml、18．7×10^5／ml,此后逐渐减少。细胞因子诱导脐血DC具有免疫刺激活性,且DC与CBMNC细胞比例为1：40时的刺激活性最佳。冻融法得到的HL-60、K562白血病细胞抗原致敏DC诱导的CTL对HL-60、K562细胞的杀伤率分别为（42．04±8．46）％和（31．25±11．07）％,与实验组比较有显著性差异（P〈0．01）。结论：加入细胞因子GM—CSF、IL-3、SCF和EPO培养2-4周的脐血MNC可分化为cD1a^＋、CD11C^＋、CD83^＋、CDw123^＋DC。冻融法得到的HL-60、K562白血病细胞抗原致敏DC,其诱导的CTL对HL-60、K562细胞具有特异的杀伤作用。脐血DC作为抗原呈递细胞在肿瘤免疫治疗上将起到重要作用。     

鳞状细胞癌抗原、糖类抗原125检测在宫颈鳞癌中的应用价值

目的探讨肿瘤标志物鳞状细胞癌抗原（SCC）与糖类抗原125（CA125）检测在宫颈鳞状细胞癌中的应用价值。方法对经临床病理确诊的85例宫颈鳞状细胞癌患者、60例宫颈上皮内瘤样病变（CIN）患者以及45例健康妇女,采用雅培微粒子免疫分析仪IMX和罗氏电化学发光分析仪E170分别检测血清SCC和CA125数值,比较分析二者改变的临床意义。结果与正常对照组及CIN组相比,宫颈鳞癌组治疗前血清SCC水平及阳性率均较高（P〈0.05）;而CA125在三组间无统计学差异（P〉0.05）。在国际妇产科联盟（Federation International of Gynecology andObstetrics,FIGO）分期中,SCC水平及阳性率随分期上升而增加（P〈0.05）;而CA125仅浓度改变有统计学差异（P〈0.05）。SCC和CA125在不同病理分级中均无显著改变（P〉0.05）。宫颈鳞癌患者治疗前后的血清SCC改变,差异有统计学意义（P〈0.05）;而血清CA125无统计学差异（P〉0.05）。结论 SCC测定可作为宫颈鳞癌诊断的辅助指标,用于病情进展、治疗效果的监测。而CA125在宫颈鳞癌诊疗中的意义不大,二者联合检测可以提高对宫颈鳞癌诊断的敏感性。     

Sonographic Findings of High‐Grade and Non–High‐Grade Ductal Carcinoma In Situ of the Breast

Objective. The purpose of this study was to differentiate between high‐grade and non–high‐grade ductal carcinoma in situ (DCIS) of the breast on sonography. Methods. From October 2003 to August 2009, 76 DCIS lesions in 73 women who underwent sonography and mammography were included in this study. Lesions were confirmed by mastectomy, breast‐conserving surgery, or surgical biopsy. Images were analyzed by 2 radiologists with consensus and were correlated with histologic grades. Results. Of the 76 lesions, 44 were classified as high‐‐grade and 32 as non–high‐grade DCIS. Fifty‐seven lesions (75.0%) were identified on sonography, which revealed a mass in 30 cases, microcalcifications in 20, ductal changes in 4, and architectural distortion in 3. All cases with false‐negative findings on sonography (n = 19) showed microcalcifications on mammography. On sonography, masses were more frequently found in non–high‐grade (62.5%) than high‐grade DCIS (22.7%; P < .01). No significant difference was seen in the sonographic features of masses between high‐grade and non–high‐grade DCIS. Microcalcifications were more common in high‐grade (43.2%) than non–high‐grade (3.1%) DCIS (P = .02). Most sonographically visible microcalcifications had associated findings such as ductal changes (n = 11), a mass (n = 7), or a hypoechoic area (n = 5). The detection rate of microcalcifications on sonography was higher in high‐grade (62.9%) than non–high‐grade DCIS (25.0%; P = .023). Conclusions. Microcalcifications with associated ductal changes (11 of 31 [35.5%]) were the most common sonographic findings in high‐grade DCIS. An irregular hypoechoic mass with an indistinct and microlobulated margin (13 of 26 [50.0%]) was the most frequent finding in non–high‐grade DCIS.     

Cell Types Obtained from the Epidural Space of Patients with Low Back Pain/Radiculopathy

Background:   We investigated if correlations exist between medical history, tissue abnormalities, and cell types retrieved from the epidural space of patients with chronic low back pain (LBP) and chronic radicular pain (RP).
Methods:   Approval was obtained from the Institutional Review Board for the Protection of Human Subjects to study 191 patients undergoing epiduroscopy. Visual inspection was performed and abnormal areas were identified. A specimen obtained from the area using a cytology brush was processed by the Thin Prep technique. Patients were divided into four groups based on the presence or absence and intensity of LBP and RP. The gender and age of the patients were recorded, as was any history of prior back surgery. Areas of tissue abnormalities were rated according to changes in vascularity and amount of fat, fibrosis, and inflammation. Stenosis was assessed from magnetic resonance imaging or computerized tomography scan images. Cytologic assessments included notations of the presence or absence of erythrocytes, leukocytes, cell groups, lipocytes, spindled cells, and large round cells.
Results:   There was a significant difference in the number of patients from whom big round cells were obtained who had a high degree of LBP compared with the number of patients who had a high degree of both LBP and RP.
Conclusions:   The findings provide a foundation for future studies of cells obtained from similar patients with the goal of furthering the understanding of the pathogenesis of LBP/RP.     

LASIK治疗近视并发症的临床观察及处理

准分子激光原位角膜磨镶术 (ＥｘｃｉｍｅｒＬａｓｅｒｉｎｓｉｔｕＫｅｒａｔｏｍｉｌｅｕｓｉｓ,ＬＡＳＩＫ)治疗近视的安全性、有效性已被大量的临床资料证实[1,2 ] 。但其手术复杂同时存在一定的并发症[3 ] 。现将本院从 1998年开展ＬＡＳＩＫ以来的病例所发生的常见并发症作一初步分析。1　资料与方法1.1　一般资料　本组病例均为临床随访半年至 1年以上的患者 ,共 896例 (162 6只眼 ) ,男性 4 0 7例 (796只眼 ) ;女性 4 89例 (830只眼 ) ;平均年龄 2 6.15 (18～5 0 )岁。屈光范围 :- 2 .0 0～ - 2 2 .0 0Ｄ ,最大散光 :-5 .0 0Ｄ。按…     

A Suppressor T Cell of the Mixed Lymphocyte Reaction Specific for the HLA-D Region in Man

The mixed lymphocyte reaction (MLR) is the proliferative response of one individual's lymphocytes cultured in the presence of another individual's lymphocytes. In man, the MLR is elicited by cell surface antigens coded for by the HLA-D gene locus. This locus is among a cluster of genes which are located on the sixth chromosome and which include genes coding for the major histocompatibility antigens HLA-A, B, and C as well as HLA-D. If the stimulator cell possesses D locus antigens not present in the responder, the lymphocytes of the latter will undergo blast transformation resulting in DNA synthesis which can be measured. A vigorous response in the MLR to allogeneic cells is the rule among healthy individuals.We describe studies of a 23-yr-old man whose lymphocytes respond normally to mitogens and soluble antigens but fail to respond to allogeneic cells in the MLR. His medical history is unremarkable except that he received thymic irradiation as an infant. HLA typing revealed that he is homozygous for HLA-A2, B12, and Cw5 as well as for the D locus antigen Dw4. When his lymphocytes were added to the responder lymphocytes of other persons homozygous for the same HLA antigens, their responses to allogeneic cells but not mitogens were suppressed by 50-95%. Their responses to a soluble antigen, tetanus toxoid, were suppressed to a lesser degree. These inhibitory effects were mediated by a relatively radioresistant thymus-derived (T) lymphocyte.Further studies of the requirements for MLR suppression revealed that only persons heterozygous or homozygous for the Dw4 antigen were inhibited by the suppressor T cell. This effect was not altered by differences in the HLA-A, B, or C antigens between the suppressor and responder. It is concluded that genes in or near the HLA-D locus code not only for antigens (primarily on bone marrow-derived (B) cells), that elicit the MLR, but also for structures on T cells, or possibly macrophages, which are recognized by MLR suppressor T cells.     

膀胱癌并前列腺增生经尿道同期电切术的临床观察

【目的】探讨膀胱癌并前列腺增生症患者同期行经尿道膀胱肿瘤加前列腺电切术治疗的可行性及疗效。【方法】对27例膀胱癌并前列腺增生患者，行同期经尿道膀胱肿瘤加前列腺电切术并随访观察疗效。【结果】随访5～60个月，5例术后复发，平均19个月，无尿道及前列腺窝种植转移。【结论】膀胱癌并前列腺增生患者同期行经尿道电切术疗效确定，未观察到有前列腺窝及尿道种植转移。     

电镜包埋后肝细胞HBVDNA原位杂交及观察

应用电镜包埋后原位分子杂交技术，对１０例人肝癌及癌周组织，分别采用ＨＢＶＤＮＡ探针进行了包埋后原位分子杂交。结果显示：６例标本呈阳性反应．其中在肝癌细胞内ＨＢＶＤＮＡ少见阳性标志．在癌周肝细胞内ＨＢＶＤＮＡ呈强弱不等的阳性反应，其分布特征，核内主要呈散在型，核外呈局部的密集型。提示ＨＢＶＤＮＡ的分布特点可能与其复制的不同时期有关．并对ＨＢＶ的致癌性进行了初步讨论．     

增殖细胞核抗原表达与鼻咽癌放射敏感性关系的研究

[目的]探讨增殖细胞核抗原(PCNA)的表达与鼻咽癌(NPC)放射敏感性的关系.[方法]采用PCNA抗体进行ABC法免疫组化染色.检测108例患者初行放疗前鼻咽癌组织中PCNA表达情况,显微镜下计数阳性细胞,计算增殖指数(PI),并将所有病例按照PI值的高低以25%为界划分两组;高PCNA表达者(HPI)>25%,低P...     

Optimisation of the Synthesis and Cell Labelling Conditions for [89Zr]Zr-oxine and [89Zr]Zr-DFO-NCS: a Direct In Vitro Comparison in Cell Types with Distinct Therapeutic Applications

Background

There is a need to better characterise cell-based therapies in preclinical models to help facilitate their translation to humans. Long-term high-resolution tracking of the cells in vivo is often impossible due to unreliable methods. Radiolabelling of cells has the advantage of being able to reveal cellular kinetics in vivo over time. This study aimed to optimise the synthesis of the radiotracers [⁸⁹Zr]Zr-oxine (8-hydroxyquinoline) and [⁸⁹Zr]Zr-DFO-NCS (p-SCN-Bn-Deferoxamine) and to perform a direct comparison of the cell labelling efficiency using these radiotracers.

Procedures

Several parameters, such as buffers, pH, labelling time and temperature, were investigated to optimise the synthesis of [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS in order to reach a radiochemical conversion (RCC) of >95 % without purification. Radio-instant thin-layer chromatography (iTLC) and radio high-performance liquid chromatography (radio-HPLC) were used to determine the RCC. Cells were labelled with [⁸⁹Zr]Zr-oxine or [⁸⁹Zr]Zr-DFO-NCS. The cellular retention of ⁸⁹Zr and the labelling impact was determined by analysing the cellular functions, such as viability, proliferation, phagocytotic ability and phenotypic immunostaining.

Results

The optimised synthesis of [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS resulted in straightforward protocols not requiring additional purification. [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS were synthesised with an average RCC of 98.4 % (n = 16) and 98.0 % (n = 13), respectively. Cell labelling efficiencies were 63.9 % (n = 35) and 70.2 % (n = 30), respectively. ⁸⁹Zr labelling neither significantly affected the cell viability (cell viability loss was in the range of 1–8 % compared to its corresponding non-labelled cells, P value > 0.05) nor the cells’ proliferation rate. The phenotype of human decidual stromal cells (hDSC) and phagocytic function of rat bone-marrow-derived macrophages (rMac) was somewhat affected by radiolabelling.

Conclusions

Our study demonstrates that [⁸⁹Zr]Zr-oxine and [⁸⁹Zr]Zr-DFO-NCS are equally effective in cell labelling. However, [⁸⁹Zr]Zr-oxine was superior to [⁸⁹Zr]Zr-DFO-NCS with regard to long-term stability, cellular retention, minimal variation between cell types and cell labelling efficiency.

     

Telavancin In Vitro Activity against a Collection of Methicillin-Resistant Staphylococcus aureus Isolates,Including Resistant Subsets,from the United States

Telavancin had MIC₅₀, MIC₉₀, and MIC₁₀₀ values of 0.03, 0.06, and 0.12 μg/ml, respectively, against methicillin-susceptible Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), and non-multidrug-resistant (non-MDR) and MDR subsets. MRSA with elevated MIC values for vancomycin (2 to 4 μg/ml) or daptomycin (1 to 2 μg/ml) had telavancin MIC₅₀ (0.06 μg/ml) values 2-fold higher than those of isolates with lower MIC results (MIC₅₀, 0.03 μg/ml). However, telavancin had MIC₉₀ and MIC₁₀₀ results of 0.06 and 0.12 μg/ml (100% susceptible), respectively, regardless of the MRSA subset.     

Evidence for Cytogenetic and Fluorescence In Situ Hybridization Risk Stratification of Newly Diagnosed Multiple Myeloma in the Era of Novel Therapies

Overall survival (OS) has improved with increasing use of novel agents in multiple myeloma (MM). However, the disease course remains highly variable, and the heterogeneity largely reflects different genetic abnormalities. We studied the impact of the Mayo risk-stratification model of MM on patient outcome in the era of novel therapies, evaluating each individual component of the model—fluorescence in situ hybridization (FISH), conventional cytogenetics (CG), and the plasma cell labeling index—that segregates patients into high- and standard-risk categories. This report consists of 290 patients with newly diagnosed MM, predominantly treated with novel agents, who were risk-stratified at diagnosis and were followed up for OS. Of these patients, 81% had received primarily thalidomide (n=50), lenalidomide (n=199), or bortezomib (n=79) as frontline or salvage therapies. Our retrospective analysis validates the currently proposed Mayo risk-stratification model (median OS, 37 months vs not reached for high- and standard-risk patients, respectively; P=.003). Although the FISH or CG test identifies a high-risk cohort with hazard ratios of 2.1 (P=.006) and 2.5 (P=.006), respectively, the plasma cell labeling index cutoff of 3% fails to independently prognosticate patient risk (hazard ratio, 1.4; P=.41). In those stratified as standard-risk by one of the 2 tests (FISH or CG), the other test appears to be of additional prognostic significance. This study validates the high-risk features defined by FISH and CG in the Mayo risk-stratification model for patients with MM predominantly treated with novel therapies based on immunomodulatory agents.CG = cytogenetics; CI = confidence interval; FISH = fluorescence in situ hybridization; HR = hazard ratio; IgH = immunoglobulin heavy chain; MM = multiple myeloma; mSMART = Mayo Stratification of Myeloma and Risk-Adapted Therapy; NR = not reached; PCLI = plasma cell labeling index; SCT = stem cell transplantationMultiple myeloma (MM) is a clonal plasma cell disorder that has witnessed considerable therapeutic advances in recent times. This progress can be attributed to improved antimyeloma therapy along with a better understanding of the tumor biology and heterogeneity.^1,2 Although a wide variation in overall survival (OS) of patients has been observed in studies analyzing the natural history of MM,³ only now are we beginning to associate the disparity in clinical outcomes with specific genetic abnormalities. Such chromosomal abnormalities are almost universally prevalent in the neoplastic plasma cells, typically occurring early in the disease process and dictating its course.⁴ Both cytogenetics (CG) and interphase fluorescence in situ hybridization (FISH) assays have played pivotal roles in the risk stratification of patients with newly diagnosed MM. Although still useful, conventional prognostic factors (β₂-microglobulin, lactate dehydrogenase, serum albumin, C-reactive protein, etc)⁵ appear to be somewhat less discerning of the outcome compared with the genetic aberrations that drive the tumor biology.^6-10The Mayo Stratification of Myeloma and Risk-Adapted Therapy (mSMART) criteria use a combination of metaphase CG, FISH, and plasma cell labeling index (PCLI; a measure of the percentage of plasma cells in the S phase of the cell cycle)¹¹ results to derive 2 composite risk categories (high-risk vs standard-risk) for prognostication of patients with newly diagnosed MM.¹² Although the initial prognostication criteria were based on the evidence predominantly garnered from patients treated with conventional chemotherapy and/or stem cell transplantation (SCT), the recommendations are periodically revised as new data emerge. Less than a decade ago, melphalan-prednisone or combination chemotherapies along with SCT were the mainstays of treatment of MM. The introduction of newer therapies, immunomodulatory drugs (thalidomide and lenalidomide), and the first-in-class proteasome inhibitor bortezomib ushered in a period of remarkable progress as the profound impact of such novel agents became evident early in the disease course.^1,13 Therefore, the Mayo prognostic model needed a formal assessment in the current era of expanded use of novel therapies. The objectives of our study were to evaluate the significance of the Mayo risk-stratification criteria since the integration of novel agents in the management of MM and to assess the independent prognostic value of each of the components (CG, FISH, and PCLI) in the model.     

Preclinical In Vitro and In Vivo Characterization of the Fully Human Monoclonal IgM Antibody KBPA101 Specific for Pseudomonas aeruginosa Serotype IATS-O11

Pseudomonas aeruginosa infection in ventilator-associated pneumonia is a serious and often life-threatening complication in intensive care unit patients, and new treatment options are needed. We used B-cell-enriched peripheral blood lymphocytes from a volunteer immunized with a P. aeruginosa O-polysaccharide-toxin A conjugate vaccine to generate human hybridoma cell lines producing monoclonal antibodies specific for individual P. aeruginosa lipopolysaccharide serotypes. The fully human monoclonal antibody secreted by one of these lines, KBPA101, is an IgM/κ antibody that binds P. aeruginosa of International Antigenic Typing System (IATS) serotype O11 with high avidity (5.81 × 10⁷ M⁻¹ ± 2.8 × 10⁷ M⁻¹) without cross-reacting with other serotypes. KBPA101 specifically opsonized the P. aeruginosa of IATS O11 serotype and mediated complement-dependent phagocytosis in vitro by the human monocyte-like cell line HL-60 at a very low concentration (half-maximal phagocytosis at 0.16 ng/ml). In vivo evaluation of KBPA101 demonstrated a dose-response relationship for protection against systemic infections in a murine burn wound sepsis model, where 70 to 100% of animals were protected against lethal challenges with P. aeruginosa at doses as low as 5 μg/animal. Furthermore, a high efficacy of KBPA101 in protection from local respiratory infections in an acute lung infection model in mice was demonstrated. Preclinical toxicology evaluation on human tissue, in rabbits, and in mice did not indicate any toxicity of KBPA101. Based on these preclinical findings, the first human clinical trials have been initiated.Pseudomonas aeruginosa is one of the leading causes of hospital-acquired (nosocomial) infections, along with coagulase-negative staphylococci, Staphylococcus aureus, and Enterococcus spp. (9, 10, 37). Bloodstream infection and pneumonia in mechanically ventilated patients are among the most frequently observed forms of nosocomial infection. Many of the commonly found bacterial strains in nosocomial infections are multidrug resistant or extensively drug resistant to most antibiotics.P. aeruginosa is a ubiquitous Gram-negative bacillus. Based on the phenotypic diversity of the O-polysaccharide moieties of surface lipopolysaccharide (LPS), P. aeruginosa is grouped into 20 unique O serotypes according to the International Antigenic Typing System (IATS) (15). Among these serotypes O11 is one of the most frequently observed ones accounting for 18 to 21% of P. aeruginosa infections (unpublished data by the authors and see also reference 28) with a very high virulence among at-risk hospital patients (42). Immunocompromised individuals in general and mechanically ventilated patients in particular are at high risk for developing pneumonia (ventilator-associated pneumonia [VAP]), for which P. aeruginosa is the most frequently detected Gram-negative bacterium (9, 37). VAP caused by P. aeruginosa is associated with significantly higher fatality rates than VAP caused by other bacterial pathogens (3, 16).The increase in antibiotic-resistant microorganisms and the paucity of new small molecule drugs for treatment of infectious diseases have renewed the interest in antibody-based anti-infective therapy (2, 33). Although high-titer antibody preparations produced by human plasma fractionation methods are well accepted for treatment of selected viral and bacterial infections, only one monoclonal antibody (MAb), namely, Palivizumab for respiratory syncytial virus, has been licensed for immunotherapy of an infectious disease thus far. However, recent advances in the generation and large-scale production of chimeric or humanized MAbs allowed the clinical development of several MAbs against viral, bacterial, and fungal diseases (29, 33, 45).IgM is the preferred isotype for complement-mediated killing and complement-dependent phagocytosis of infectious bacteria due to its pentameric form and its ability for effective complement activation. Furthermore, polysaccharides (including LPS) are T-cell-independent antigens, and antibodies induced in response to them are mostly of the IgM isotype. However, recombinant expression of pentameric IgM has not been achieved routinely thus far. There have been many attempts to generate human MAbs of different isotypes against various antigens of P. aeruginosa, such as LPS, alginate, or PcrV (reviewed by Doring and Pier [8]). Most of these MAbs demonstrated protection in animal models (18, 25, 30, 39, 41, 47, 48), and some were subsequently tested in clinical settings (12, 27, 38), but none of these MAbs have been licensed for routine use in humans thus far.We describe here the generation and preclinical characterization of a fully human IgM/κ MAb termed KBPA101, directed against the LPS O polysaccharide of serotype O11 of P. aeruginosa. KBPA101 was generated by immortalizing human B lymphocytes and selected for high effector function. KBPA101 demonstrated high specificity and efficacy in vitro and in vivo and was already tested in a clinical phase 1 study (23).     

介入治疗联合适形放射治疗原发性肝细胞癌的疗效分析

目的：探讨经导管肝动脉灌注化疗栓塞（TACE）与TACE联合三维适形放射治疗（TACE＋3DCRT）原发性肝细胞癌（HCC）的疗效。方法：随机分成两组的HCC患者,均不能手术、门脉无癌栓、无远处脏器转移。50例患者行单纯TACE2次（TACE组）。40例患者在TACE2次后第四周再行三维适形放射治疗（TACE＋3DCRT组）,放疗采用6MV-X射线,4～6Gy／次,隔日一次,3次／周,总剂量45～60Gy。TACE采用碘化油、顺铂、5-氟尿嘧啶、表阿霉素及明胶海绵。结果：TACE组与TACE＋3DCRT组近期疗效有效率（完全缓解＋部分缓解）分别为68．0％（34／50）、87．5％（35／40）,1年生存率分别为78．0％（39／50）、95．0％（38／40）,两组差异均有统计学意义（P〈0．05）。TACE的治疗反应主要为栓塞综合征,3DCRT有7．5％（3／40）发生急性放射性肝炎,经对症、保肝处理后缓解。两组患者均能耐受。结论：TACE＋3DCRT治疗不能手术、门脉无癌栓、无远处脏器转移的HCC的疗效较单纯TACE治疗HCC的疗效好。