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M Stow

Publications and source records attributed to M Stow.

5 recordsLinked to original sources
PubMed2001

Analysis of cellular adhesion by microarray expression profiling.

Microarrays of oligonucleotides or cDNAs can be used to establish the expression profiles of numerous genes in a single experiment. We have established a microarray platform to identify genes in a number of different pathological conditions, particularly those with an inflammation component. This platform utilised the output of an eosinophil sequencing project in which 1069 sequences were identified that were not represented in the public domain. An eosinophil model cell line, AML14.3D10, was used to investigate cell adhesion. The transcription profile of adhered and non-adhered AML 14.3D10 cells was shown to be both technically and biologically reproducible. A number of genes were found differentially expressed in the adhered vs. non-adhered populations. In the adhered population, the expression of these genes was restricted compared to brain, lung, kidney and especially bone marrow. However, the differentially regulated genes were not among those genes most restricted to eosinophils. We discuss the implications of transcription profiling on gene annotation and its potential utility for the identification of targets for drug intervention.

Base Sequence↗
PubMed1993

Molecular mechanisms resulting in pathogenic anti-mouse erythrocyte antibodies in New Zealand black mice.

The New Zealand black (NZB) mouse strain is genetically predisposed to develop, at approximately 6 months of age, a spontaneous and severe autoimmune anaemia caused by production of pathogenic anti-mouse erythrocyte autoantibodies. In order to investigate the molecular mechanisms which lead to anti-mouse erythrocyte autoantibody production we have generated eight anti-mouse erythrocyte MoAbs producing hybridomas from splenocytes of 9- and 12-month-old NZB with spontaneous autoimmune anaemia. IgG2a was the predominant isotype, while IgM, IgG1 and IgG2b were each produced by one hybridoma cell line. All anti-mouse erythrocyte MoAbs were characterized for their antigen specificities. None of the MoAbs cross-reacted with ss- or dsDNA or with other species' erythrocytes, with the exception of one MoAb which cross-reacted with rat erythrocytes. None of the eight hybridomas was demonstrated to express surface or cytoplasmic CD5, suggesting that they derived from CD5- B lymphocytes. All hybridomas when implanted intraperitoneally into BALB/c mice caused anaemia. In order to define the genetic basis and investigate the molecular mechanisms resulting in pathogenic anti-mouse erythrocyte autoantibody production, the pattern of immunoglobulin variable region gene use has been studied. Five of the eight MoAbs whose IgVH genes were sequenced all have functionally rearranged genes from the VH J558 gene family. There is evidence for somatic point mutations in the complementarity-determining regions (CDR) of the IgVH genes in all of these five MoAbs when compared with the closest known germline gene. We suggest that these nucleotide sequence changes are likely to reflect selection by an antigen-driven mechanism. Furthermore, the data indicate that pathogenic anti-mouse erythrocytes are not derived from 'natural' autoantibodies.

Amino Acid Sequence↗
PubMed1993

Anti-mouse red blood cell monoclonal antibodies use functionally rearranged genes from the VH J558 family and are derived from the CD5- B-lymphocyte subpopulation.

The NZB mouse strain is genetically predisposed to develop, at approximately 6 months of age, a spontaneous and severe autoimmune anaemia caused by the production of pathogenic anti-mouse red blood cell (MRBC) autoantibodies. Although it is believed that the predisposition to autoimmune anaemia is multigenic in nature, the main pathogenic mechanism is attributed to anti-MRBC autoantibodies. We have generated eight anti-MRBC monoclonal antibody (mAb)-producing hybridomas derived from splenocytes of 9- and 12-month-old NZB mice with spontaneous autoimmune anaemia to dissect the molecular and cellular mechanisms resulting in the production of these pathogenic antibodies. The predominant immunoglobulin isotype was IgG2a, produced by five out of eight hybridomas (63%), while IgM, IgG1 and IgG2b were each produced by one hybridoma cell line (12%). Antigen specificity analysis of all eight hybridomas revealed that antibodies from seven out of eight hybridomas were monospecific for MRBC antigen(s). Only one hybridoma (clone 4-16-1) cross-reacted with rat RBC. None of the hybridomas produced antibodies reactive with single- or double-stranded DNA (ss- or dsDNA). Surface and cytoplasmic staining for the CD5 antigen revealed that none of the hybridomas was derived from CD5+ B lymphocytes. All hybridomas cause anaemia when implanted intraperitoneally into normal BALB/c mice. Molecular studies of five of the eight anti-MRBC mAb reveal that all use functionally rearranged genes from the VH J558 gene family. Three of these five mAb used FL16.1 DH genes while one had a CDR3 that resulted from a fusion between two DH genes (SP2.3 and SP2.2) from the SP family.

Anemia↗