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Article: Chronic hepatitis C and genotyping: the clinical significance of determining HCV genotypes.

Hnatyszyn, H James

2005 Volume 10, Issue 1, Page(s) 1–11

Abstract: Chronic hepatitis C, attributed to infection with hepatitis C virus (HCV), is a global health problem. The overall prevalence of viral hepatitis worldwide is estimated to be 3-5% with over 175 million people infected with HCV. Clinically, HCV can ... ...

Abstract	Chronic hepatitis C, attributed to infection with hepatitis C virus (HCV), is a global health problem. The overall prevalence of viral hepatitis worldwide is estimated to be 3-5% with over 175 million people infected with HCV. Clinically, HCV can establish a persistent, chronic infection contributing to progressive liver disease, including cirrhosis and hepatocellular carcinoma (HCC), requiring intensive treatment regimens, possible liver transplantation and long-term care. Due to the chronic nature of HCV infection and the tremendous burden on healthcare resources, clinicians and laboratorians have looked for key epidemiological, pathological and viral characteristics that may provide insight into disease progression, severity and response to therapy to permit the administration of effective therapeutic regimens as well as long-term management of infected individuals. Determination of viral genotype has been identified as one parameter that could provide direction in the clinical management of patients with chronic HCV infections. The following review provides background on determination of HCV genotypes and the relevance of viral genome characterization in the current clinical setting.
MeSH term(s)	Algorithms ; Antiviral Agents/therapeutic use ; Genotype ; Hepacivirus/genetics ; Hepatitis C, Chronic/drug therapy ; Hepatitis C, Chronic/epidemiology ; Hepatitis C, Chronic/virology ; Humans ; Interferon Type I/therapeutic use ; Recombinant Proteins
Chemical Substances	Antiviral Agents ; Interferon Type I ; Recombinant Proteins
Language	English
Publishing date	2005
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1339842-8
ISSN	1359-6535
ISSN	1359-6535
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Article: Chronic hepatitis B and viral genotype: the clinical significance of determining HBV genotypes.

Guettouche, Toumy / Hnatyszyn, H James

Antiviral therapy

2005 Volume 10, Issue 5, Page(s) 593–604

Abstract: The global health challenge posed by the hepatitis B virus (HBV) centres around the widespread distribution and the serious complications as a result of persistent infection with the virus. As with other chronic diseases mediated by pathogens such as ... ...

Abstract	The global health challenge posed by the hepatitis B virus (HBV) centres around the widespread distribution and the serious complications as a result of persistent infection with the virus. As with other chronic diseases mediated by pathogens such as human immunodeficiency virus (HIV) and hepatitis C virus (HCV), clinicians are searching for epidemiological, pathological and viral characteristics of HBV infection that may lead to more effective management of patients with chronic infection. Unlike HCV, the role of HBV genotype in disease progression, severity, response to therapy and drug resistance is still under investigation and just beginning to be clarified. This review examines the potential role of HBV genotype determination in the clinic with emphasis on how this genetic information may used to provide effective management for the treatment of patients with chronic hepatitis B.
MeSH term(s)	Algorithms ; Antiviral Agents/therapeutic use ; Disease Progression ; Genotype ; Global Health ; Hepatitis B virus/genetics ; Hepatitis B, Chronic/diagnosis ; Hepatitis B, Chronic/drug therapy ; Hepatitis B, Chronic/virology ; Humans ; Treatment Outcome
Chemical Substances	Antiviral Agents
Language	English
Publishing date	2005
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	1339842-8
ISSN	1359-6535
ISSN	1359-6535
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Article ; Online: Quantitative and qualitative characterization of Two PD-L1 clones: SP263 and E1L3N.

Smith, Jacquelyn / Robida, Mark D / Acosta, Krista / Vennapusa, Bharathi / Mistry, Amita / Martin, Greg / Yates, Alton / Hnatyszyn, H James

Diagnostic pathology

2016 Volume 11, Issue 1, Page(s) 44

Abstract: ... staining. H scores were used for membrane scoring whereas percent positivity was used for tumor-associated ... with the SP263 assay. Dynamic range was demonstrated by the SP263 assay having well-distributed H scores ... while the E1L3N assay had a significantly higher proportion of cases clustered in the lowest H score bins ...

Abstract	Background: Programmed Death Ligand 1 (PD-L1) is an immune modulating protein expressed on the surface of various inflammatory cells, including T Cells, B Cells, dendritic cells, and macrophages. PD-L1 represents an important diagnostic target; expression of PD-L1 on the surface of tumor cells, or within tumor-associated immune cells, is an important predictor of likely response to targeted therapies. In this study, we describe the optimization of immunohistochemistry (IHC) assays using two PD-L1 antibodies (SP263 and E1L3N) and compare the performance of the optimized assays. Methods: Fully automated immunohistochemical assays were optimized for the VENTANA PD-L1 (SP263) Rabbit Monoclonal Antibody and the PD-L1 (E1L3N®) XP® Rabbit mAb using instruments and detection chemistries from Ventana Medical Systems, Inc. ("SP263 assay" and "E1L3N assay," respectively). Tissue microarrays (TMAs) containing formalin fixed paraffin embedded (FFPE) non-small cell lung cancer (NSCLC) cases were used for the optimization and comparison staining. H scores were used for membrane scoring whereas percent positivity was used for tumor-associated immune cell scoring. Results: One-hundred NSCLC TMA case cores each stained with the SP263 and E1L3N assays were evaluated by two pathologists in a blinded study. Analysis of these specimens showed that the SP263 assay was more sensitive and had a wider dynamic range than the E1L3N assay. For sensitivity, many cases were found to be negative for membrane staining with the E1L3N assay, but had measurable staining with the SP263 assay. Dynamic range was demonstrated by the SP263 assay having well-distributed H scores while the E1L3N assay had a significantly higher proportion of cases clustered in the lowest H score bins. For tumor-associated immune cell staining, the two assays identified similar amounts of cells staining in each case, but the SP263 assay gave overall darker staining. Conclusion: Since PD-L1 status is important for targeted therapies, having a specific and accurate diagnostic test is crucial for identifying patients who could benefit from these treatments. Due to its staining intensity, scoring range, and pathologist preference, the SP263 IHC assay has been deemed superior to the E1L3N IHC assay. Future clinical utility remains to be determined.
MeSH term(s)	Animals ; Antibodies, Monoclonal/immunology ; B7-H1 Antigen/genetics ; B7-H1 Antigen/immunology ; B7-H1 Antigen/metabolism ; Biomarkers, Tumor/metabolism ; Carcinoma, Non-Small-Cell Lung/diagnosis ; Carcinoma, Non-Small-Cell Lung/metabolism ; Humans ; Immunohistochemistry ; Lung Neoplasms/diagnosis ; Lung Neoplasms/metabolism ; Rabbits ; Staining and Labeling ; Tissue Array Analysis
Chemical Substances	Antibodies, Monoclonal ; B7-H1 Antigen ; Biomarkers, Tumor ; CD274 protein, human
Language	English
Publishing date	2016-05-18
Publishing country	England
Document type	Comparative Study ; Evaluation Study ; Journal Article
ZDB-ID	2210518-9
ISSN	1746-1596 ; 1746-1596
ISSN (online)	1746-1596
ISSN	1746-1596
DOI	10.1186/s13000-016-0494-2
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Article ; Online: The role of SATB1 in breast cancer pathogenesis.

Iorns, Elizabeth / Hnatyszyn, H James / Seo, Pearl / Clarke, Jennifer / Ward, Toby / Lippman, Marc

Journal of the National Cancer Institute

2010 Volume 102, Issue 16, Page(s) 1284–1296

Abstract: Background: SATB1 has been previously proposed as a key protein that controls the development and progression of breast cancer. We explored the potential of the SATB1 protein as a therapeutic target and prognostic marker for human breast cancer.: ... ...

Abstract	Background: SATB1 has been previously proposed as a key protein that controls the development and progression of breast cancer. We explored the potential of the SATB1 protein as a therapeutic target and prognostic marker for human breast cancer. Methods: We used aggressive (MDA-MB-231 and BT549) and nonaggressive (SKBR3 and MCF7) breast cancer cell lines to investigate the potential of SATB1 as a therapeutic target. SATB1 mRNA expression was silenced in aggressive cells by use of short hairpin RNAs against SATB1. SATB1 was overexpressed in nonaggressive cells by use of SATB1 expression vectors. We assessed the effect of modifying SATB1 expression on the transformed phenotype by examining anchorage-independent cell proliferation, acinar morphology on matrigel, and migration by wound healing in cultured cells. We examined tumor formation and metastasis, respectively, by use of orthotopic mammary fat pad and tail vein xenograft mouse models (mice were used in groups of six, and in total, 96 mice were used). SATB1 mRNA expression was compared with outcome for patients with primary breast cancer from six previous microarray studies that included a total of 1170 patients. All statistical tests were two-sided. Results: The transformed phenotype was not suppressed by SATB1 silencing in aggressive cells and was not enhanced by ectopic expression of SATB1 in nonaggressive cells. Modifying SATB1 expression did not alter anchorage-independent cell proliferation, invasive acinar morphology, or cell migration in cultured cells and did not affect tumor formation or metastasis in xenograft mouse models. In addition, SATB1 expression was not associated with decreased overall survival of patients with primary breast cancer in six previous independent microarray studies (overall odds ratio = 0.80, 95% confidence interval = 0.62 to 1.03, P = .10). Conclusion: In contrast to previous studies, we found that SATB1 expression did not promote breast cancer progression and was not associated with breast cancer outcome.
MeSH term(s)	Animals ; Biomarkers, Tumor/genetics ; Biomarkers, Tumor/metabolism ; Breast Neoplasms/metabolism ; Cell Line, Tumor ; Collagen ; Drug Combinations ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Immunoblotting ; Laminin ; Matrix Attachment Region Binding Proteins/genetics ; Matrix Attachment Region Binding Proteins/metabolism ; Mice ; Mice, Nude ; Oligonucleotide Array Sequence Analysis ; Predictive Value of Tests ; Prognosis ; Proteoglycans ; RNA Interference ; RNA, Messenger/metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors/metabolism ; Transplantation, Heterologous ; Up-Regulation ; Wound Healing
Chemical Substances	Biomarkers, Tumor ; Drug Combinations ; Laminin ; Matrix Attachment Region Binding Proteins ; Proteoglycans ; RNA, Messenger ; SATB1 protein, human ; SATB2 protein, human ; Transcription Factors ; matrigel (119978-18-6) ; Collagen (9007-34-5)
Language	English
Publishing date	2010-08-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2992-0
ISSN	1460-2105 ; 0027-8874 ; 0198-0157
ISSN (online)	1460-2105
ISSN	0027-8874 ; 0198-0157
DOI	10.1093/jnci/djq243
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Article: HPV in HIV-Infected Women: Implications for Primary Prevention.

McKenzie, Nathalie Dauphin / Kobetz, Erin N / Ganjei-Azar, Parvin / Rosa-Cunha, Isabella / Potter, JoNell E / Morishita, Atsushi / Lucci, Joseph A / Guettouche, Toumy / Hnatyszyn, James H / Koru-Sengul, Tulay

Frontiers in oncology

2014 Volume 4, Page(s) 179

Abstract: Background: There is growing evidence that human immunodeficiency virus (HIV)-infected women might have a different human papillomavirus (HPV) type distribution in cervical dysplasia specimens as compared to the general population. This has implications ...

Abstract	Background: There is growing evidence that human immunodeficiency virus (HIV)-infected women might have a different human papillomavirus (HPV) type distribution in cervical dysplasia specimens as compared to the general population. This has implications for primary prevention. Objective: We aimed to obtain preliminary data on the HPV genotypes prevalent in histological samples of HIV-infected women with cervical intraepithelial neoplasia (CIN) 3/CIS of the cervix in Miami, FL, USA. Methods: Retrospective data were collected on HIV-infected women referred to the University of Miami-Jackson Memorial Hospital colposcopy clinic between years 2000 and 2008. The histology slides of CIN 3/CIS biopsies underwent pathological review and sections were cut from these archived specimens for HPV DNA extraction. HPV genotyping was then performed using the GeneSquare™ HPV genotyping assay. We report on our first set of 23 samples. Results: Eight high-risk HPV types were detected. Types in decreasing order of frequency were 16, 35, 45, 52, 59, 31, 58, and 56. Most cases had multiple infections. HPV type 16 was the most common (45%) followed by HPV-35 and -45 with equal frequency (40%). No samples contained HPV-18. Conclusion: Our preliminary results suggest that cervical dysplasia specimens of HIV-infected women more likely (55%) contain non-16 and -18 high-risk HPV types. We show that this held true for histologically confirmed severe dysplasia and carcinoma-in situ. Epidemiological studies guide vaccine development, therefore HPV type prevalence in CIS and invasive cervical cancer among HIV-infected women should be more rigorously explored to ensure that this highly vulnerable population receives appropriate primary prevention.
Language	English
Publishing date	2014-08-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2649216-7
ISSN	2234-943X
ISSN	2234-943X
DOI	10.3389/fonc.2014.00179
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Article: Rapid and specific detection of Mycobacterium tuberculosis from acid-fast bacillus smear-positive respiratory specimens and BacT/ALERT MP culture bottles by using fluorogenic probes and real-time PCR.

Miller, Nancimae / Cleary, Tim / Kraus, Günter / Young, Andrea K / Spruill, Gina / Hnatyszyn, H James

Journal of clinical microbiology

2001 Volume 40, Issue 11, Page(s) 4143–4147

Abstract: ... within 1 h of having a DNA sample available for testing. ...

Abstract	A real-time PCR assay using the LightCycler (LC) instrument for the specific identification of Mycobacterium tuberculosis complex (MTB) was employed to detect organisms in 135 acid-fast bacillus (AFB) smear-positive respiratory specimens and in 232 BacT/ALERT MP (MP) culture bottles of respiratory specimens. The LC PCR assay was directed at the amplification of the internal transcribed spacer region of the Mycobacterium genome with real-time detection using fluorescence resonance energy transfer probes specific for MTB. The results from the respiratory specimens were compared to those from the Amplicor M. tuberculosis PCR test. Specimens from MP culture bottles were analyzed by Accuprobe and conventional identification methods. MTB was cultured from 105 (77.7%) respiratory AFB smear-positive specimens; 103 of these samples were positive by LC PCR and Amplicor PCR. Two samples negative in the LC assay contained rare numbers of organisms; both were positive in the Amplicor assay. Two separate samples negative by Amplicor PCR contained low and moderate numbers of AFB, respectively, and both of these were positive in the LC assay. There were 30 AFB smear-positive respiratory specimens that grew mycobacteria other than tuberculosis (MOTT), and all tested negative in both assays. Of the 231 MP culture bottles, 114 cultures were positive for MTB and all were positive by the LC assay. The remaining 117 culture bottles were negative in the LC assay and grew various MOTT. This real-time MTB assay is sensitive and specific; a result was available within 1 h of having a DNA sample available for testing.
MeSH term(s)	Bacterial Typing Techniques ; Culture Media ; Fluorescence Resonance Energy Transfer ; Fluorescent Dyes ; Humans ; Mycobacterium tuberculosis/genetics ; Mycobacterium tuberculosis/growth & development ; Mycobacterium tuberculosis/isolation & purification ; Polymerase Chain Reaction/methods ; Reagent Kits, Diagnostic ; Respiratory System/microbiology ; Sensitivity and Specificity ; Specimen Handling/methods ; Temperature ; Time Factors ; Tuberculosis, Pulmonary/microbiology
Chemical Substances	Culture Media ; Fluorescent Dyes ; Reagent Kits, Diagnostic
Language	English
Publishing date	2001-10-13
Publishing country	United States
Document type	Evaluation Study ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	390499-4
ISSN	1098-660X ; 0095-1137
ISSN (online)	1098-660X
ISSN	0095-1137
DOI	10.1128/JCM.40.11.4143-4147.2002
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Article ; Online: Whole genome in vivo RNAi screening identifies the leukemia inhibitory factor receptor as a novel breast tumor suppressor.

Iorns, Elizabeth / Ward, Toby M / Dean, Sonja / Jegg, Anna / Thomas, Dafydd / Murugaesu, Nirupa / Sims, David / Mitsopoulos, Costas / Fenwick, Kerry / Kozarewa, Iwanka / Naceur-Lombarelli, Cristina / Zvelebil, Marketa / Isacke, Clare M / Lord, Christopher J / Ashworth, Alan / Hnatyszyn, H James / Pegram, Mark / Lippman, Marc

Breast cancer research and treatment

2012 Volume 135, Issue 1, Page(s) 79–91

Abstract: Cancer is caused by mutations in oncogenes and tumor suppressor genes, resulting in the deregulation of processes fundamental to the normal behavior of cells. The identification and characterization of oncogenes and tumor suppressors has led to new ... ...

Abstract	Cancer is caused by mutations in oncogenes and tumor suppressor genes, resulting in the deregulation of processes fundamental to the normal behavior of cells. The identification and characterization of oncogenes and tumor suppressors has led to new treatment strategies that have significantly improved cancer outcome. The advent of next generation sequencing has allowed the elucidation of the fine structure of cancer genomes, however, the identification of pathogenic changes is complicated by the inherent genomic instability of cancer cells. Therefore, functional approaches for the identification of novel genes involved in the initiation and development of tumors are critical. Here we report the first whole human genome in vivo RNA interference screen to identify functionally important tumor suppressor genes. Using our novel approach, we identify previously validated tumor suppressor genes including TP53 and MNT, as well as several novel candidate tumor suppressor genes including leukemia inhibitory factor receptor (LIFR). We show that LIFR is a key novel tumor suppressor, whose deregulation may drive the transformation of a significant proportion of human breast cancers. These results demonstrate the power of genome wide in vivo RNAi screens as a method for identifying novel genes regulating tumorigenesis.
MeSH term(s)	Basic Helix-Loop-Helix Leucine Zipper Transcription Factors/genetics ; Breast Neoplasms/drug therapy ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Cell Line, Tumor ; Female ; Genes, Tumor Suppressor ; Genes, p53 ; Genome, Human ; High-Throughput Nucleotide Sequencing ; Humans ; Leukemia Inhibitory Factor Receptor alpha Subunit/genetics ; RNA Interference ; RNA, Small Interfering ; Repressor Proteins/genetics ; Tumor Suppressor Protein p53/genetics
Chemical Substances	Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; LIFR protein, human ; Leukemia Inhibitory Factor Receptor alpha Subunit ; MNT protein, human ; RNA, Small Interfering ; Repressor Proteins ; TP53 protein, human ; Tumor Suppressor Protein p53
Language	English
Publishing date	2012-04-26
Publishing country	Netherlands
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	604563-7
ISSN	1573-7217 ; 0167-6806
ISSN (online)	1573-7217
ISSN	0167-6806
DOI	10.1007/s10549-012-2068-7
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Zs.A 1655: Show issues

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Article: Cross-clade inhibition of HIV-1 replication and cytopathology by using RNase P-associated external guide sequences.

Kraus, Gunter / Geffin, Rebeca / Spruill, Gina / Young, Andrea K / Seivright, Rachel / Cardona, Diana / Burzawa, Jennifer / Hnatyszyn, H James

Proceedings of the National Academy of Sciences of the United States of America

2002 Volume 99, Issue 6, Page(s) 3406–3411

Abstract: RNase P complexes have been proposed as a novel RNA-based gene interference strategy to inhibit gene expression in human malignancies and infectious diseases. This approach is based on the sequence-specific design of an external guide sequence (EGS) RNA ... ...

Abstract	RNase P complexes have been proposed as a novel RNA-based gene interference strategy to inhibit gene expression in human malignancies and infectious diseases. This approach is based on the sequence-specific design of an external guide sequence (EGS) RNA molecule that can specifically hybridize to almost any complementary target mRNA and facilitate its cleavage by the RNase P enzyme component. We designed a truncated RNase P-associated EGS molecule to specifically recognize the U5 region of HIV-1 mRNA and mediate cleavage of hybridized mRNA by the RNase P enzyme. Genes encoding for this U5-EGS (560) molecule, as well as a U5 EGS (560D) antisense control, were cloned into retroviral plasmids and transferred into a CD4(+) T cell line. Transfected cells were exposed to increasing concentrations of HIV-1 clinical isolates from clades A, B, C, and F. Heterogeneous cultures of CD4(+) T cells expressing the U5 EGS (560) molecule were observed to maintain CD4 levels, were devoid of cytopathology, and did not produce HIV p24 gag antigen through 30 days after exposure to all HIV-1 clades at a multiplicity of infection of 0.01. Identical cells expressing the U5 EGS (560D) antisense control molecule underwent a loss of CD4 expression, produced elevated levels of HIV-1, and formed large syncytia similar to untreated cells. When the viral inoculum was increased at the time of exposure (multiplicity of infection = 0.05), the inhibitory effect of the U5 EGS (560) molecule was overwhelmed, but viral-mediated cytopathology and particle production were delayed compared with control cell populations. Viral replication and cytopathology associated with infection of multiple HIV-1 clades can be effectively inhibited in CD4(+) cells expressing the RNase P-associated U5 EGS (560) molecule.
MeSH term(s)	CD4-Positive T-Lymphocytes/metabolism ; CD4-Positive T-Lymphocytes/virology ; Cell Line ; Cytopathogenic Effect, Viral ; DNA, Complementary/genetics ; DNA, Viral/biosynthesis ; DNA, Viral/genetics ; Endoribonucleases/biosynthesis ; Endoribonucleases/genetics ; Endoribonucleases/metabolism ; Flow Cytometry ; HIV Core Protein p24/biosynthesis ; HIV Core Protein p24/genetics ; HIV-1/classification ; HIV-1/genetics ; HIV-1/metabolism ; HIV-1/physiology ; Humans ; Plasmids/genetics ; Proviruses/genetics ; RNA, Catalytic/biosynthesis ; RNA, Catalytic/genetics ; RNA, Catalytic/metabolism ; Retroviridae/genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonuclease P ; Substrate Specificity ; Transfection ; Virus Replication ; RNA, Small Untranslated
Chemical Substances	DNA, Complementary ; DNA, Viral ; HIV Core Protein p24 ; RNA, Catalytic ; Endoribonucleases (EC 3.1.-) ; RPP14 protein, human (EC 3.1.26.5) ; Ribonuclease P (EC 3.1.26.5) ; RNA, Small Untranslated
Language	English
Publishing date	2002-06-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.052651199
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Article: Multicenter evaluation of the VERSANT hepatitis B virus DNA 3.0 assay.

Yao, Joseph D C / Beld, Marcel G H M / Oon, Lynette Lin Ean / Sherlock, Christopher H / Germer, Jeffrey / Menting, Sandra / Se Thoe, Su Yun / Merrick, Linda / Ziermann, Rainer / Surtihadi, Johan / Hnatyszyn, H James

Journal of clinical microbiology

2003 Volume 42, Issue 2, Page(s) 800–806

Abstract: The VERSANT hepatitis B virus (HBV) 3.0 Assay (branched DNA [bDNA]) (referred to herein as VERSANT 3.0) was evaluated at four external sites for analytical sensitivity, specificity, reproducibility, linearity of quantification, and limits of detection. ... ...

Abstract	The VERSANT hepatitis B virus (HBV) 3.0 Assay (branched DNA [bDNA]) (referred to herein as VERSANT 3.0) was evaluated at four external sites for analytical sensitivity, specificity, reproducibility, linearity of quantification, and limits of detection. In addition, each of the test evaluation sites provided HBV DNA-positive clinical samples that were previously analyzed by one of three commercially available HBV DNA quantitative tests: Digene Hybrid Capture II HBV DNA Test (Digene); VERSANT HBV DNA 1.0 Assay (bDNA) (VERSANT 1.0); and COBAS AMPLICOR HBV Monitor Test (COBAS AMPLICOR). These samples were reexamined using VERSANT 3.0. The results from these studies showed that VERSANT 3.0 has high specificity (99.3%), excellent reproducibility (between-run coefficient of variation [CV] = 1.6 to 9.4%; within-run CV = 6.5 to 20.7%), and a broad linear range of quantification (2.0 x 10(3) to 1.0 x 10(8) HBV DNA copies/ml) that facilitate the monitoring of HBV DNA levels at diagnosis and throughout the course of treatment. In general, correlation was very good between results obtained from clinical samples analyzed by VERSANT 3.0 and the comparative HBV DNA quantitative assays (VERSANT 1.0, R(2) = 0.900; Digene, R(2) = 0.985; COBAS AMPLICOR, R(2) = 0.771). The greatest differences in comparative quantitation occurred at HBV DNA levels approaching the limits of the dynamic ranges for the comparative assays. The performance characteristics of the new VERSANT 3.0 assay demonstrated that it provides a reliable and robust method for routinely monitoring serum HBV DNA levels in assessing disease activity and determining response to antiviral treatment.
MeSH term(s)	DNA, Viral/blood ; DNA, Viral/genetics ; Genetic Techniques ; Hepatitis B/diagnosis ; Hepatitis B virus/genetics ; Humans ; Reproducibility of Results ; Sensitivity and Specificity
Chemical Substances	DNA, Viral
Language	English
Publishing date	2003-11-17
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Multicenter Study
ZDB-ID	390499-4
ISSN	1098-660X ; 0095-1137
ISSN (online)	1098-660X
ISSN	0095-1137
DOI	10.1128/JCM.42.2.800-806.2004
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Article: Induced dendritic cell differentiation of chronic myeloid leukemia blasts is associated with down-regulation of BCR-ABL.

Lindner, Inna / Kharfan-Dabaja, Mohamed A / Ayala, Ernesto / Kolonias, Despina / Carlson, Louise M / Beazer-Barclay, Yasmin / Scherf, Uwe / Hnatyszyn, James H / Lee, Kelvin P

Journal of immunology (Baltimore, Md. : 1950)

2003 Volume 171, Issue 4, Page(s) 1780–1791

Abstract: Although differentiation of leukemic blasts to dendritic cells (DC) has promise in vaccine strategies, the mechanisms underlying this differentiation and the differences between leukemia and normal progenitor-derived DC are largely undescribed. In the ... ...

Abstract	Although differentiation of leukemic blasts to dendritic cells (DC) has promise in vaccine strategies, the mechanisms underlying this differentiation and the differences between leukemia and normal progenitor-derived DC are largely undescribed. In the case of chronic myeloid leukemia (CML), understanding the relationship between the induction of DC differentiation and the expression of the BCR-ABL oncogene has direct relevance to CML biology as well as the development of new therapeutic approaches. We now report that direct activation of protein kinase C (PKC) by the phorbol ester PMA in the BCR-ABL(+) CML cell line K562 and primary CML blasts induced nonterminal differentiation into cells with typical DC morphology (cytoplasmic dendrites), characteristic surface markers (MHC class I, MHC class II, CD86, CD40), chemokine and transcription factor expression, and ability to stimulate T cell proliferation (equivalent to normal monocyte-derived DC). PKC-induced differentiation was associated with down-regulation of BCR-ABL mRNA expression, protein levels, and kinase activity. This down-regulation appeared to be signaled through the mitogen-activated protein kinase pathway. Therefore, PKC-driven differentiation of CML blasts into DC-like cells suggests a potentially novel strategy to down-regulate BCR-ABL activity, yet raises the possibility that CML-derived DC vaccines will be less effective in presenting leukemia-specific Ags.
MeSH term(s)	Blast Crisis/genetics ; Blast Crisis/immunology ; Blast Crisis/pathology ; Calcium/metabolism ; Calcium/physiology ; Cell Differentiation/drug effects ; Cell Differentiation/genetics ; Cell Differentiation/immunology ; Cell Division/drug effects ; Cell Division/immunology ; Cell Lineage/genetics ; Cell Lineage/immunology ; Cells, Cultured ; Cytokines/pharmacology ; Dendritic Cells/enzymology ; Dendritic Cells/pathology ; Down-Regulation/genetics ; Enzyme Activation/drug effects ; Enzyme Activation/genetics ; Fusion Proteins, bcr-abl/antagonists & inhibitors ; Fusion Proteins, bcr-abl/biosynthesis ; Fusion Proteins, bcr-abl/genetics ; Gene Expression Regulation, Neoplastic/drug effects ; Gene Expression Regulation, Neoplastic/immunology ; Genes, abl/immunology ; Humans ; Intracellular Fluid/metabolism ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/immunology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology ; Protein Kinase C/metabolism ; Protein Kinase C/physiology ; Protein-Tyrosine Kinases ; Proto-Oncogene Proteins/antagonists & inhibitors ; Proto-Oncogene Proteins/biosynthesis ; Proto-Oncogene Proteins/genetics ; Proto-Oncogene Proteins c-bcr ; Signal Transduction/genetics ; Signal Transduction/immunology ; Tetradecanoylphorbol Acetate/pharmacology ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha/pharmacology
Chemical Substances	Cytokines ; Proto-Oncogene Proteins ; Tumor Necrosis Factor-alpha ; Protein-Tyrosine Kinases (EC 2.7.10.1) ; Fusion Proteins, bcr-abl (EC 2.7.10.2) ; BCR protein, human (EC 2.7.11.1) ; Proto-Oncogene Proteins c-bcr (EC 2.7.11.1) ; Protein Kinase C (EC 2.7.11.13) ; Tetradecanoylphorbol Acetate (NI40JAQ945) ; Calcium (SY7Q814VUP)
Language	English
Publishing date	2003-07-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	3056-9
ISSN	1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
ISSN (online)	1550-6606
ISSN	0022-1767 ; 1048-3233 ; 1047-7381
DOI	10.4049/jimmunol.171.4.1780
Database	MEDical Literature Analysis and Retrieval System OnLINE

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