• Cell-Penetrating Peptides

      Howl, John; Jones, Sarah (2015)
      The multi-domain architecture of many human proteins provides a structural basis for the physical maintenance of interactomes, or networks of protein-protein interactions (PPIs), that are so obviously crucial to cellular functions. Moreover, the structural and electrostatic complementarity provided by PPI interfaces, predominantly located on protein surfaces, is a fundamental component of signal transduction events that are known to be compromised in human diseases including many cancers.The pharmacokinetic advantages provided by cell-penetrating peptides (CPPs) are entirely compatible with the development of intrinsically permeable agents capable of modulating intracellular PPIs. Thus, the term bioportide is a useful descriptor of numerous bioactive CPPs that are distinct from the more usual inert CPP vectors. Herein we illustrate a generic strategy, predominantly centered upon the identification of cationic peptides derived from helical protein domains, which offers a reliable platform to identify bioportides capable of modulating intracellular signal transduction events. In addition, we describe robust methodologies to determine the precise intracellular distribution of fluorescent bioportides and present assays routinely employed to screen for the detrimental pharmacodynamic properties often exhibited by both CPPs and bioportides; namely adverse cytotoxicity and the receptor-independent stimulation of mast cell secretion.
    • Case study of Felty’s Syndrome

      Nelson, Paul N.; Bowman, S.J. (Oxford: Blackwell Publishing, 2001)
      This book: is an introductory level text on the biological principles of human disease. The book is aimed at medical students in degree courses in biomedical science. The book fuses the biological (physiological and biochemical) processes which underlie the clinical manifestations of disease. As such, it brings together material which is conventionally dealt with by several books. The authors have covered the fundamentals of each topic in a readable manner, which should encourage students to develop a fuller understanding, where necessary, by reference to more comprehensive texts.
    • Generating monoclonal antibody probes and techniques for characterizing and localizing reactivity to antigenic determinants

      Nelson, Paul N. (Oxford: Oxford University Press, 2001)
      This book: Epitope Mapping covers all the major methods for the identification and definition of epitopes. The Pepscan assay is used to define B cell epitopes and makes use of synthetic peptides but can only be used if the amino acid sequence is known. It can be adapted for the delineation of both helper T cells and cytotoxic T cells. The identification of combined B and T cell epitopes can also be achieved using synthetic peptides. There are other methodologies for analysing for cytotxic T cell epitopes such as the purification of antigens presented by MHC class I molecules and expression cloning. Site directed mutagenesis is also a powerful tool in epitope mapping and can be used to evaluate the role of single amino acids in immune complex formation. Protein footprinting makes use of monoclonal antibodies produced by hybridoma technology and relies on the fact that the epitope is protected from cleavage when bound as an antibody-antigen complex. It is only useful for small antigens. Other monoclonal antibody assays such as enzyme linked immunosorbent assay and haemaglutination and slot-blotting may also be used in epitope mapping. Random phage display libraries bring together the genetic and amino acid peptide sequence and can be screened with antibody and the resulting peptide DNA sequenced to confirm the amino acid sequence of a specific eptiope. Investigation of carbohydrates can also be useful to eptitope mapping as deglycosylation can lead to loss of antigenic activity. Epitopes are important to the pharmaceutical industry and wherever appropriate, pharmaceutical applications of the methods described are included. For each method there is a description of the technology, protocols, trouble-shooting, and advice on when to use the method. This book will therefore be invaluable to any researcher involved in epitope mapping.
    • Monoclonal antibodies: The tools of the trade within biomedical science

      Nelson, Paul N.; Astley, S.J.; Warren, Phil (John Wiley & Sons, Ltd., 2003)
      This book: The latest edition of this highly successful text, covers the major advances in the methods used in cellular and molecular pathology. In recent years, knowledge of the molecular organization of the cell has led to the development of powerful new techniques that bring greater accuracy and objectives to the diagnosis, prognosis and management of many diseases and to the study of pathological states. This book describes the latest molecular techniques available for the analysis of diseases. In particular it includes new techniques using fluorescent dyes, DNA microarrays, protein chemistry, and mass spectrometry. It also incorporates information from the Human Genome Project, and the new disciplines of genomics and proteomics, where relevant to pathology. Color plates are a new feature of this edition, illustrating the advances in fluorescence labeling of cells.
    • Generation and characterization of monoclonal antibodies to the neural crest.

      Shakil, T.; Richardson, M. K.; Waldron, E.E.; Conde, Gillian; Wood, S.; Bland, Y.; Reynolds, Gary; Murray, Paul G.; Nelson, Paul N. (New Rochelle (NY): Mary Ann Liebert, Inc., 2001)
      The generation of monoclonal antibodies (MAbs) specific for quail neural crest may provide valuable tools for studying the differentiation of embryonic precursor cells. Unfortunately, relatively few antibodies are available because of the difficulty in obtaining sufficient cells for in vivo immunization strategies. We have overcome this problem by using intrasplenic immunization with formaldehyde-fixed cells harvested from neural crest cultures. In addition, booster injections of cultured whole-embryo cells were administered intraperitoneally. Following two fusions, a total of 18 hybridomas were generated with antibody reactivity to the cytoplasm of neural crest cells. Furthermore, 32 were reactive against both somite (a noncrest mesodermal control) and crest cultures, whilst 15 were not reactive. Out of those hybridomas reactive with neural crest, six designated 160D, 164D, OE, 12E, 120E and 124E were further characterized. Interestingly MAb supernatants OE, 12E, 120E, and 124E exhibited reactivity against some but not all neural crest cells suggesting that they might recognise subpopulations. Immunoglobulin isotyping of supernatants revealed that 4 (160D, 164D, OE, and 120E) were IgM and 2 (12E and 124E) were IgG(2b). On assessing their reactivity against human tissue sections, all six hybridoma supernatants cross-reacted with neuroendocrine cells within appendix, colon and rectum. These MAbs could provide novel reagents for the understanding of neural crest development.
    • Effect of material deprivation on Epstein-Barr virus infection in Hodgkin's disease in the West Midlands.

      Flavell, Joanne R.; Constandinou, C.; Lowe, D.; Scott, K.; Newey, C.; Evans, D.; Dutton, A.; Simmons, S.; Smith, Richard; Crocker, John; Young, Lawrence S.; Murray, Paul G. (nature.com, 1999)
      We have used Townsend scores from postcode data to compare levels of material deprivation and Epstein-Barr virus (EBV)-positivity for 223 patients diagnosed with Hodgkin's disease (HD) in the period 1981-1997. The presence of EBV in HD tumours was determined using in situ hybridization to target the abundantly expressed EBV early RNAs. EBV was detected in the malignant Hodgkin and Reed-Sternberg cells in 47/223 HD cases (21%). There was found to be a tendency for higher Townsend scores (indicative of higher levels of material deprivation) in EBV-positive HD patients, but this association was not statistically significant. When various subgroups of patients from the study were examined separately the indication of higher Townsend scores in EBV-positive patients was found to be more marked for patients with mixed cellularity disease (P = 0.09) and for females (P = 0.03). The results of this study suggest that differences in the level of material deprivation are important in determining the likelihood of EBV-positive HD in the UK, particularly for certain subgroups of patients. It is not known what specific socioeconomic factors are responsible for these differences, although alterations in the timing or rate of primary EBV infection, or decline in the level of EBV-specific immunity, may be important. (Cancer Research UK)
    • Acid phosphatases.

      Bull, H.; Murray, Paul G.; Thomas, David G.; Fraser, A. M.; Nelson, Paul N. (BMJ Publishing Group Ltd., 2002)
      Acid phosphatases (APs) are a family of enzymes that are widespread in nature, and can be found in many animal and plant species. Mystery surrounds the precise functional role of these molecular facilitators, despite much research. Yet, paradoxically, human APs have had considerable impact as tools of clinical investigation and intervention. One particular example is tartrate resistant acid phosphatase, which is detected in the serum in raised amounts accompanying pathological bone resorption. This article seeks to explore the identity and diversity of APs, and to demonstrate the relation between APs, human disease, and clinical diagnosis.
    • Human endogenous retrovirus HERV-K10 implicated in rheumatoid arthritis and systemic lupus erythematosus: potential pathological triggers?

      Nelson, Paul N.; Shaw, M.; Roden, Denise A.; Freimanis, Graham L.; Nevill, Alan M.; Rylance, Paul (Czech Republic: Palacky University, Olomouc: Faculty of Medicine and Dentistry, 2006)
      Human endogenous retroviruses (HERVs) are a group of integrated RNA viruses within our human genome. Whilst many are regarded as defective, a number possess the potential to generate retroviral products. Indeed HERVs such as those belonging to the HERV-K family produce retroviral particles in the teratocarcinoma cell line GH and the breast cancer cell line T47D. It has been argued that some retroelements may be beneficial to the human host, perhaps conferring a selective advantage, whereas others may be harmful. Furthermore certain HERVs might be involved in the pathogenesis of autoimmune diseases. The precise mechanisms in diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) may include molecular mimicry and superantigen motifs that evoke and augment unwarranted immune responses. The precise mechanisms in diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) may include molecular mimicry and superantigen motifs that evoke and augment unwarranted immune responses. In the RA joint, tissue destruction is evident over time with recruitment of lymphoid and other cells plus the presence of rheumatoid factor that exhibits increased affinity and change in isotype; evidence of an antigen-driven immune response. The precise trigger of course, remains unknown although certain HERVs have been implicated. In a previous study we found evidence for increased expression of HERV-K10 mRNA in patients with RA. Here we have extended this work by investigating the serological expression to HERV-K10 in patients with RA, SLE, osteoarthritis, normals and other inflammatory disease groups. The study utilised a novel peptide ELISA immunoassay using segments of HERV-K10 identified through bioinformatic analysis. In particular, biotinylation of peptides was necessary for serological discrimination between patients. Overall a significant difference (p<0.05) was found for RA patients in terms of antibody activity to HERV-K10. There was also an increased level of antibodies to HERV-K10 in patients with renal lupus although this was below the level of significance. It is possible that HERV-K10 could act as a trigger in RA/SLE through regions of similarity to host proteins. In this case, the immune response to HERV-K10 could lead to collateral damage and pathogenesis of disease.
    • Comparison of Antigenic Regions Identified on IgG1Fc Using Bioinformatics vs Pepscan Analysis

      Nelson, Paul N.; Westwood, Olwyn M. R.; Freimanis, Graham L.; Roden, Denise A.; Sissaoui, Samir; Rylance, Paul; Hay, Frank C. (Libertas Academica Press, 2008)
      Epitope mapping allowed the location of antigenic determinants on a protein macromolecule to be identified. In particular, pepscan techniques that utilize a series of overlapping peptides, help detect key amino acid residues that are important in antibody recognition and binding. In a previous study, we employed 15-mer peptides spanning the entire length of IgG1Fc to ascertain successfully the target epitopes of isotypic/allotypic monoclonal reagents. As an extension to this work we have used these peptides to evaluate the location of epitope targets of five IgM rheumatoid factor antibodies (RFAbs). Overall, 2 antibodies, RFAb TS2 and TS1, detected a similar epitope within the CH3 domain (360-KNQVSLTCLVKGFYP-374), whilst 1 (RFAb SJ1) recognised an epitope in the CH2 domain (294- EQYNSTYRVVSVLTV-308). In contrast, 2 RFAbs, PRSJ2 and PRTS1 detected four and five epitopes respectively within the Fc region. RFAb PRSJ2 recognised epitopes detected by RFAB TS2 and TS1 but also further epitopes in the CH2 domain (256-TPEVTCVVVDVSHED-270) and CH3 domain (418-QQGNVFSCSVMHEAL-432). Similarly, RFAb PRTS1 detected all four epitopes plus a fifth in the CH3 domain (382-ESNGQPENNYKTTPP-396). In essence there was a consensus of target epitopes identified by these rheumatoid factor antibodies. Interestingly, two epitopes (256–270, CH2 domain and 360–374, CH3 domain) were novel in that they had not been identified in previous pepscan studies. The other epitopes recognised, either overlapped or were immediately adjacent to previous epitopes detected by poly/monoclonal rheumatoid factor antibodies. Molecular modelling (PCImdad) of IgG1Fc showed that all five epitopes were exposed and surface accessible for antibody interaction. In addition, a bioinformatics analysis of the Fc region using ExPASy was employed to identify key antigenic determinants. This ‘in silico’ approach may provide a means of determining key regions without the need to develop overlapping peptides spanning the entire length of a macromolecule.
    • Reactivity and isotype profiling of monoclonal antibodies using multiple antigenic peptides.

      Waldron, E.E.; Murray, Paul G.; Kolar, Zdenek; Young, Lawrence S.; Brown, C.; Reynolds, Gary; Baumforth, Karl R. N.; Toomey, S.; Astley, S.J.; Perera, Shantha; Nelson, Paul N. (New Rochelle (NY): Mary Ann Liebert, Inc., 2002)
      The characterisation of monoclonal antibodies (MAbs) is essential for the development of assay systems particularly where antigens have been developed using synthetic peptides. Indeed some peptide-carrier conjugates fail to induce immune responses and may not generate antibodies that bind to native protein. As an alternative to peptide-carrier conjugates, multiple antigenic peptides (MAPs) have been used for immunization strategies, but with little regard to the characteristics of the MAbs produced. In this study, we used 3 MAPs of Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) to immunise BALB/c mice. Overall, the polyclonal antibody responses from tail bleeds showed that MAPs evoked B-cell responses. However, on screening 144 hybridomas, 24 MAb supernatants exhibited weak to moderate reactivity in enzyme-linked immunosorbant assay (ELISA) and against cell cytospin preparations (B95.8 and AG876 LCL), respectively. Isotype profiling of hybridoma supernatants also showed that 11 out of 24 were IgM. Further characterization of 6 MAbs in Western blotting showed reactivity to recombinant LMP1 and only one MAb (B28D) showed weak reactivity to the malignant cells (Hodgkin/Reed-Sternberg; HRS cells) of an EBV+ Hodgkin's lymphoma using paraffin-embedded tissue. It is probable that these MAPs failed to augment T-cell help and contributed to the production of low affinity (IgM) antibodies. These observations may be of importance to future immunization strategies, where MAPs are used in the production of monoclonal reagents.
    • Demystified...recombinant antibodies.

      Smith, K.A.; Nelson, Paul N.; Warren, Phil; Astley, S.J.; Murray, Paul G.; Greenman, J. (BMJ Publishing Group Ltd., 2004)
      Recombinant antibodies are important tools for biomedical research and are increasingly being used as clinical diagnostic/therapeutic reagents. In this article, a background to humanized antibodies is given, together with details of the generation of antibody fragments--for example, single chain Fv fragments. Phage antibody fragments are fast becoming popular and can be generated by simple established methods of affinity enrichment from libraries derived from immune cells. Phage display methodology can also be used for the affinity enrichment of existing antibody fragments to provide a reagent with a higher affinity. Here, phage antibodies are demystified to provide a greater understanding of the potential of these reagents and to engage clinicians and biomedical scientists alike to think about potential applications in pathology and clinical settings.
    • Characterization of anti-myosin monoclonal antibodies.

      Nelson, Paul N.; Astley, S.J.; Roden, Denise A.; Waldron, E.E.; Baig, K.M.; Caforio, A.L.; Koutedakis, Yiannis; Perera, Shantha; Spry, C. (New Rochelle (NY): Mary Ann Liebert, Inc., 2005)
      The characterization of monoclonal antibodies (MAbs) with regard to reactivity and specificity is important for the successful application as a molecular probe and/or diagnostic reagent. Furthermore, it is recognized that some monoclonal reagents perform well in some assay systems but not others. In this study, the reactivity profiles of two anti-myosin MAbs (H1 and DH2, raised against human cardiac myosin) were evaluated in enzyme-linked immunosorbent assay (ELISA), slot-blotting, and immunocytochemistry. Both antibodies performed well in slot-blotting against myosin heavy chain preparations from cardiac and skeletal muscle and from non-human sources. In general, MAb H1 demonstrated strong to moderate reactivity in all assay systems, whilst MAb DH2 faired poorly in ELISA. MAb H1 also showed reactivity to synthetic peptides of myosin, one of which possessed a motif (ERRDA, single amino acid code) that was found in other human and nonhuman myosin protein sequences that could explain its cross-reactive profile. Intriguingly, this motif was found on viral and other pathogenic agents associated with myocarditis. Hence, it is speculated that this region could give some credence to the mechanism of molecular mimicry associated with some cardiac diseases. Overall, MAb H1 may serve as a useful probe of myosin structure.
    • Rheumatoid factors: what's new?

      Westwood, Olwyn M. R.; Nelson, Paul N.; Hay, Frank C. (Oxford: Oxford Journals, 2006)
      Rheumatoid arthritis (RA) is a classic example of an autoimmune disorder, with chronic inflammation of the synovial membrane, and deterioration of cartilage and bone in the affected joints. The resultant pain, loss of function and permanent disability are also associated with increased morbidity and mortality.
    • Does microalbuminuria predict illness severity in critically ill patients on the intensive care unit? A systematic review.

      Gopal, Shameer; Carr, Bryan; Nelson, Paul N. (Lippincott Williams & Wilkins, 2006)
      CONTEXT: Studies assessing the accuracy of microalbuminuria to predict illness severity on the intensive care unit have produced inconsistent results. OBJECTIVE: To determine the diagnostic accuracy of microalbuminuria to predict illness severity in critically ill patients on the intensive care unit. DATA SOURCE: MEDLINE (1951 to September 2004) and EMBASE (1980 to September 2004) electronic databases were searched for relevant studies. Reference lists of all abstracts were manually searched to identify studies not included in the electronic database. STUDY SELECTION: Studies that prospectively evaluated the accuracy of microalbuminuria to predict illness severity and/or mortality probability in adult patients on the intensive care unit were selected. DATA EXTRACTION: We included nine studies in the review. Data to evaluate methodological quality and results were abstracted. DATA SYNTHESIS: The methodological quality of a number of studies was poor. Significant heterogeneity in the design and conduct of the studies circumvented the data being subjected to meta-analysis. Studies also differed in the timing of the index test, in the methods of quantifying microalbuminuria, and in the cutoff values used. CONCLUSIONS: This descriptive analysis reveals that microalbuminuria may hold promise as a predictor of illness severity and mortality on the intensive care unit. However, future epidemiologic studies need to be conducted to determine the optimal timing as well as the threshold reference value for the urine albumin creatinine ratio in the adult intensive care unit population. Thereafter, multiple-center prospective epidemiologic studies must be conducted to confirm and validate the findings of these preliminary studies. Future studies should conform to the Standards for Reporting of Diagnostic Accuracy checklist in terms of study design, conduct, and reporting. Presently there is no evidence to warrant the use of this tool on the intensive care unit. (Lippincott Williams & Wilkins, Inc.)
    • Molecular pathology of brain tumours - how will molecular and cell biology contribute to improved outcomes in patients with malignant brain tumours?

      Darling, John L. (Czech Republic: Palacky University, Olomouc: Faculty of Medicine and Dentistry, 2006)
      Although in comparison to breast, lung and colon cancer, the brain is a relatively uncommon site for the development of cancer, the brain is the tenth most common site for the development of cancer in men and about the twelfth in women. This translates to about 6,000 individuals in the UK developing a primary malignant brain tumour every year. Cancer of the brain develops in two distinct age groups, although the types of tumour that develop in these two age groups differ markedly. There is a peak of incidence in the first decade of life, and brain tumours rank with leukaemia as a leading cause of cancer death in children. These tumours tend to be indolent low-grade astrocytomas or highly malignant primitive neuroectodermal tumours like medulloblastoma. However, the vast majority of brain tumours occur with increasing frequency in the sixth, seventh and eight decade of life and they are the second fastest growing cause of cancer death among those over 65. These tumours tend to be malignant astrocytomas particularly the most malignant variety, glioblastoma multiforme (GBM). Unlike lung cancer or malignant melanoma there is no strong evidence of an environmental carcinogen associated with the development of these tumours and no change in behaviour reduces risk.
    • South Asian ethnicity and material deprivation increase the risk of Epstein-Barr virus infection in childhood Hodgkin's disease.

      Flavell, K.J.; Biddulph, J.P.; Powell, J.E.; Parkes, S.E.; Redfern, D.; Weinreb, M.; Nelson, Paul N.; Mann, J.R.; Young, Lawrence S.; Murray, Paul G. (Nature Publishing Group, 2001)
      In order to further define the factors associated with the observed variations in the Epstein-Barr virus-positive rate in childhood Hodgkin's disease, we have studied the effect of material deprivation (measured by the Townsend score) and ethnic origin on the frequency of Epstein-Barr virus-positivity in 55 cases of childhood Hodgkin's disease, diagnosed between 1981 and 1999, from a multi-ethnic region of the United Kingdom. Epstein-Barr virus status was determined by immunohistochemistry for the Epstein-Barr virus-encoded latent membrane protein-1. 62% of cases were Epstein-Barr virus-positive. Ethnic group was the strongest predictor of Epstein-Barr virus-positivity, with South Asians having a more than 20-fold risk of being Epstein-Barr virus-positive compared with non-South Asians. An increased risk was still present after adjusting for deprivation. Townsend scores were significantly higher (indicating more deprivation) in the Epstein-Barr virus-positive group, particularly in males. The relative risk of Epstein-Barr virus-positivity showed a gradient with increasing Townsend score; the risk being 7-times higher in the most deprived quartile compared with the least deprived group. Although the association between Townsend score and Epstein-Barr virus-positivity was reduced after adjusting for ethnic group, the risk of Epstein-Barr virus-positivity was still 3-times higher in the most deprived compared with the least deprived quartile. In addition, cases having 2 or more siblings were 5-times as likely to be Epstein-Barr virus-positive as those from smaller families. These results provide the first evidence of a strong association between Epstein-Barr virus-positive Hodgkin's disease and South Asian children from the United Kingdom. In addition, deprivation may increase the likelihood of Epstein-Barr virus-positive disease independently of ethnicity.
    • Interleukin 6 expression by Hodgkin/Reed-Sternberg cells is associated with the presence of 'B' symptoms and failure to achieve complete remission in patients with advanced Hodgkin's disease.

      Reynolds, Gary; Billingham, Lucinda; Gray, Laura; Flavell, Joanne R.; Najafipour, Sohrab; Crocker, John; Nelson, Paul N.; Young, Lawrence S.; Murray, Paul G. (Blackwell Synergy, 2002)
      Interleukin 6 (IL-6) is a potent immunomodulatory cytokine that has pathogenic and prognostic significance in a number of disorders. Previous studies in Hodgkin's disease (HD) have demonstrated the association between elevated serum levels of IL-6 and unfavourable prognosis, including advanced stage and the presence of 'B' symptoms and with reduced survival. Although IL-6 expression has been demonstrated in both the malignant Hodgkin/Reed-Sternberg (HRS) cells and in the various non-malignant cells present in HD biopsies, a relationship between expression of IL-6 by the tumour and outcome measures has not been established. The study group comprised of 97 patients with advanced HD who were recruited to two related clinical trials. IL-6 expression was determined on paraffin-wax sections of biopsy material by means of an immunohistochemical assay. Of the 97 patients, 27 (28%) showed staining for IL-6 in HRS cells. IL-6 expression by HRS cells was significantly correlated with a decreased likelihood of achieving a complete response to chemotherapy (P = 0.02) and with an increased prevalence of 'B' symptoms (P = 0.04). IL-6 expression by HRS cells was not associated with Epstein-Barr virus status (P = 0.57). In summary, the results suggest that IL-6 expression by HRS cells may contribute to the presence of 'B' symptoms and to a decreased likelihood to achieve a complete remission in HD patients.
    • The potential role of human endogenous retrovirus K10 in the pathogenesis of rheumatoid arthritis: a preliminary study.

      Ejtehadi, H. Davari; Freimanis, Graham L.; Ali, H.A.; Bowman, S.J.; Alavi, A.; Axford, J.; Callaghan, R.; Nelson, Paul N. (BMJ Publishing, 2006)
      OBJECTIVE: To examine whether human endogenous retrovirus K10 is associated with autoimmune rheumatic disease. DESIGN: A novel multiplex reverse transcription polymerase chain reaction (RT-PCR) system was developed to investigate HERV-K10 mRNA expression in patients with rheumatoid arthritis. METHODS: 40 patients with rheumatoid arthritis, 17 with osteoarthritis, and 27 healthy individuals were recruited and total RNA was extracted from peripheral blood mononuclear cells (PBMCs) and analysed using multiplex RT-PCR for the level of HERV-K10 gag mRNA expression. Southern blot and DNA sequencing confirmed the authenticity of the PCR products. RESULTS: Using the histidyl tRNA synthetase (HtRNAS) gene as a housekeeping gene in the optimised multiplex RT-PCR, a significantly higher level of HERV-K10 gag mRNA expression was found in rheumatoid arthritis than in osteoarthritis (p = 0.01) or in the healthy controls (p = 0.02). CONCLUSION: There is enhanced mRNA expression of the HERV-K10 gag region in rheumatoid arthritis compared with osteoarthritis or healthy controls. This could contribute to the pathogenesis of rheumatoid arthritis.
    • Role of sexual behavior in the acquisition of asymptomatic Epstein-Barr virus infection: a longitudinal study.

      Woodman, Ciaran; Collins, Stuart; Vavrusova, Nicol; Rao, Ankit; Middeldorp, Jaap; Kolar, Zdenek; Kumari, Angela; Nelson, Paul N.; Young, Lawrence S.; Murray, Paul G. (Lippincott Williams & Wilkins, 2005)
      BACKGROUND: The natural history of Epstein-Barr virus (EBV) infection is poorly defined. We report the prevalence and subsequent incidence of EBV infection in a cohort of sexually active young women and explore the social and sexual determinants of incident infections. METHODS: The study population was drawn from a cohort of young women, who were recruited for a longitudinal study of risk factors for early cervical neoplasia. A case-control analysis, nested within the cohort of 45 women for whom the first EBV sample tested was EBV-negative and who had further follow-up, was undertaken. EBV serostatus was determined in serum with a synthetic peptide-based enzyme-linked immunosorbent assay; EBV DNA was measured in cervical smears with the use of quantitative polymerase chain reaction. RESULTS: Of 1023 women 15-19 years of age included in this analysis, 978 (95.6%) tested positive for antibodies to EBV in their first serum sample. Of 45 women who tested negative, 22 subsequently acquired an asymptomatic EBV infection; the median time to seroconversion was 25 months (range, 1-60 months), and the median age at seroconversion was 18 years (range, 16-21 years). The risk of seroconversion increased with increasing number of sexual partners [compared with 1 partner, odds ratio (OR) was 1.28 for 2 partners and 2.23 for 3 or more; chiTREND 5.02; df 1; P < 0.05] and was greatest when a new sexual partner had been acquired in the 2 years before seroconversion (OR 4.78; chi 4.62; df 1; P < 0.05). EBV DNA was detected in 9 of 14 women who seroconverted and who also provided cervical samples. CONCLUSIONS: In susceptible young women, the acquisition of EBV infection is associated with their sexual behavior.
    • The prevalence and clinical significance of autoantibodies to plasminogen activator inhibitor 1 in systemic lupus erythematosus.

      Bates, Ruth L.; Payne, Sarah J.; Drury, S.L.; Nelson, Paul N.; Isenberg, D.A.; Murphy, John J.; Frampton, Geoffrey (SAGE Publications, 2003)
      We have recently described the novel autoantigen plasminogen activator inhibitor (PAI-1) in systemic lupus erythematosus (SLE). The aim of this study was to determine the prevalence and clinical significance of anti-PAI-1 autoantibodies in patients with SLE. Autoantibodies to recombinant PAI-1 were measured in retrospective sera of 48 lupus patients by immunoassay in order to assess their clinical significance. This showed that 71% of sera from 48 lupus patients had significantly elevated anti-PAI-1 autoantibodies as compared with normal control subjects (P < 0.0001). There was a weak but significant (P < 0.043) correlation with anti-dsDNA autoantibodies. In longitudinal studies, autoantibodies against PAI-1 correlated with clinical parameters measured by the BILAG disease activity index including global clinical score. Our study demonstrates the high frequency of novel autoantibodies to PAI-1 in patients with lupus. The serial clinical correlations with anti-PAI-1 autoantibodies also support the hypothesis that these autoantibodies may play a pathogenic role in lupus.