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41.
Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus 总被引:7,自引:0,他引:7
Berry JD Jones S Drebot MA Andonov A Sabara M Yuan XY Weingartl H Fernando L Marszal P Gren J Nicolas B Andonova M Ranada F Gubbins MJ Ball TB Kitching P Li Y Kabani A Plummer F 《Journal of virological methods》2004,120(1):87-96
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report, the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented, based upon their specificity, binding requirements, and biological activity. Initial screening by ELISA, using highly purified virus as the coating antigen, resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb, F26G15, is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients, but not normal human serum, is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests, studies of SARS-CoV pathogenesis and vaccine development. 相似文献
42.
43.
Berge-Lefranc JL; Jay P; Massacrier A; Cau P; Mattei MG; Bauer S; Marsollier C; Berta P; Fontes M 《Human molecular genetics》1996,5(10):1637-1641
While constructing a cDNA library of human embryos, we have isolated a
clone homologous to jumonji, a mouse gene required for neural tube
formation. We have determined the complete coding sequence of the human
homologue (JMJ) and deduced the amino acid sequence of the putative
protein. We show here that human and mouse jumonji putative proteins are
homologous and present 90% identity. During human embryogenesis, JMJ mRNAs
are predominantly expressed in neurons and particularly in dorsal root
ganglion cells. They are also expressed in neurons of human adult cerebral
cortex. In view of these observations, we propose JMJ as a candidate gene
for developmental defects of the central nervous system in the human. The
human JMJ gene maps at position 6p24-6p23.
相似文献
44.
An investigation into the relationship between salivary cortisol,stress, anxiety and depression 总被引:3,自引:0,他引:3
Vedhara K Miles J Bennett P Plummer S Tallon D Brooks E Gale L Munnoch K Schreiber-Kounine C Fowler C Lightman S Sammon A Rayter Z Farndon J 《Biological psychology》2003,62(2):89-96
This study examined the relationship between indices of self-reported emotional distress and absolute versus change in cortisol levels. Fifty-four women attending a diagnostic breast clinic completed scales measuring stress, anxiety and depression and provided five saliva samples over the course of a single day for the measurement of cortisol. No significant relationships were evident between absolute cortisol levels and the distress measures. Analysis of the change in cortisol levels revealed a non-linear interaction effect between stress and anxiety and time of day. There was a non-linear relation between time of day and cortisol levels, but the extent of the non-linearity was dependent upon levels of stress and anxiety, not depression. A relationship was apparent between indices of distress and change in cortisol levels, but not absolute levels of the hormone. 相似文献
45.
Casey Graham; Plummer Sarah; Hoeltge Gerald; Scanlon David; Fasching Clare; Stanbridge Eric J. 《Human molecular genetics》1993,2(11):1921-1927
Rearrangements or deletions of chromosome 17 are the most frequentlyobserved genetic changes identified in breast tumors. Molecularanalyses suggest that in addition to the p53 gene on 17p13.1there may be at least three other tumor suppressor genes onchromosome 17 involved in breast cancer. Regions of loss ofheterozygosity (LOH) identified on 17p13.3 and 17q12-qter occurfrequently in breast tumors, and the BRCA-1 gene has been mappedto 17q21 by genetic linkage analysis. Here we provide biologicalevidence for the presence of a growth suppressor gene(s) onchromosome 17 that results In the In vitro growth suppressionof the p53 wild-type MCF 7 breast cancer cell line. We haveIntroduced a normal chromosome 17 into MCF 7 cells by microcellmediatedchromosome transfer (MMCT), and demonstrate that cells growtharrest before 10 to 12 population doublings. In contrast, theintroduction of a normal chromosome 13 had no effect upon growthof these cells either In vitro or In vivo. These data providedirect functional evidence for the presence of a growth suppressorgene(s) on chromosome 17, which is not p53, and which may representone of several gene(s) that play a critical role in the developmentof breast cancer. 相似文献
46.
How important is the 'quality' of the cytotoxic T lymphocyte (CTL) response in protection against HIV infection? 总被引:7,自引:0,他引:7
Rowland-Jones SL Pinheiro S Kaul R Hansasuta P Gillespie G Dong T Plummer FA Bwayo JB Fidler S Weber J McMichael A Appay V 《Immunology letters》2001,79(1-2):15-20
Cytotoxic T lymphocyte (CTL) responses have been associated with protection from HIV-1 infection in people with a high degree of exposure to HIV and who show no serological evidence of HIV infection (HEPS, highly exposed persistently seronegative). However, it remains unclear how protective CTL responses could apparently develop in a minority of people, whilst the great majority of HIV-infected people make strong CTL responses yet progress to AIDS and death. In this paper we review the data which supports the hypothesis that the quality of the T-cell response, rather than its magnitude, may be an important factor that merits further investigation. 相似文献
47.
How to use Chlamydia antibody testing in subfertility patients 总被引:1,自引:9,他引:1
Screening for tubal factor subfertility by means of Chlamydia antibody
testing (CAT) was introduced into the initial work-up of subfertile couples
several years ago. The results reported, however, are heterogeneous, and no
uniformity exists in cut-off levels of titres, or in definitions of tubal
factor subfertility. We performed a prospective cohort study to evaluate
the implications of varying the definitions of tubal pathology and of
modifying the cut-off levels on the clinical impact of CAT in predicting
tubal factor subfertility. In 227 consecutive patients who attended our
fertility clinic, the Chlamydia IgG antibody titre was determined and
related to tuboperitoneal abnormalities at laparoscopy as a reference
standard. According to received operating characteristic (ROC) curve
analysis, a titre of 16 is the optimum cut-off level. Increasing the
cut-off level improves specificity and positive likelihood ratio (LR+), at
the expense of sensitivity and negative LR (LR-). Changing the definition
of tubal factor subfertility from unspecified tuboperitoneal abnormalities
into extensive adhesions and/or bilateral distal tubal occlusion improves
LR+, LR- and kappa significantly. We conclude that CAT is more accurate in
predicting severe distal tubal pathology than unspecified tuboperitoneal
abnormalities. Although from a statistical point of view a titre of 16 is
the optimum cut-off level, from a clinical point of view 32 or 64 may be
preferable, depending on the aim of screening and the inception cohort.
相似文献
48.
Pal L; Leykin L; Schifren JL; Isaacson KB; Chang YC; Nikruil N; Chen Z; Toth TL 《Human reproduction (Oxford, England)》1998,13(7):1837-1840
A case series of eight cycles of in-vitro fertilization (IVF) in five women
diagnosed with malignant disorders is presented. These patients chose to
defer definitive treatment for a chance for preservation of potential
fertility. The response of these patients to ovarian stimulation, and the
outcome, was compared with 17 IVF cycles in 12 age- matched patients with
isolated tubal infertility. An apparent adverse influence of malignant
disease on the quality and behaviour of oocytes was observed. Despite a
comparable total number of oocytes per cycle in the two groups, a
significantly reduced percentage of mature oocytes was retrieved per cycle
from patients with malignant diseases. The oocytes from patients with
malignant disorders were of a poorer quality and exhibited a significantly
impaired fertilization rate compared to the controls. We propose that
neoplastic processes, irrespective of the site or cell of origin, may have
a detrimental impact on the biology of oocytes, an effect akin to that seen
on spermatozoa in men with certain malignancies.
相似文献
49.
50.
Distribution of mutations in the PEX gene in families with X-linked hypophosphataemic rickets (HYP) 总被引:8,自引:0,他引:8
Rowe PS; Oudet CL; Francis F; Sinding C; Pannetier S; Econs MJ; Strom TM; Meitinger T; Garabedian M; David A; Macher MA; Questiaux E; Popowska E; Pronicka E; Read AP; Mokrzycki A; Glorieux FH; Drezner MK; Hanauer A; Lehrach H; Goulding JN; O'Riordan JL 《Human molecular genetics》1997,6(4):539-549
Mutations in the PEX gene at Xp22.1 (phosphate-regulating gene with
homologies to endopeptidases, on the X-chromosome), are responsible for
X-linked hypophosphataemic rickets (HYP). Homology of PEX to the M13 family
of Zn2+ metallopeptidases which include neprilysin (NEP) as prototype, has
raised important questions regarding PEX function at the molecular level.
The aim of this study was to analyse 99 HYP families for PEX gene
mutations, and to correlate predicted changes in the protein structure with
Zn2+ metallopeptidase gene function. Primers flanking 22 characterised
exons were used to amplify DNA by PCR, and SSCP was then used to screen for
mutations. Deletions, insertions, nonsense mutations, stop codons and
splice mutations occurred in 83% of families screened for in all 22 exons,
and 51% of a separate set of families screened in 17 PEX gene exons.
Missense mutations in four regions of the gene were informative regarding
function, with one mutation in the Zn2+-binding site predicted to alter
substrate enzyme interaction and catalysis. Computer analysis of the
remaining mutations predicted changes in secondary structure,
N-glycosylation, protein phosphorylation and catalytic site molecular
structure. The wide range of mutations that align with regions required for
protease activity in NEP suggests that PEX also functions as a protease,
and may act by processing factor(s) involved in bone mineral metabolism.
相似文献