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Titolo:
IMMUNODETECTION OF HISTONE EPITOPES CORRELATES WITH EARLY STAGES OF APOPTOSIS IN ACTIVATED HUMAN PERIPHERAL T-LYMPHOCYTES
Autore:
ZUNINO SJ; SINGH MK; BASS J; PICKER LJ;
Indirizzi:
UNIV TEXAS,SW MED CTR,DEPT PATHOL,LAB MOL PATHOL,5323 HARRY HINES BLVD DALLAS TX 75235 UNIV TEXAS,SW MED CTR,DEPT PATHOL,LAB MOL PATHOL DALLAS TX 75235
Titolo Testata:
The American journal of pathology
fascicolo: 2, volume: 149, anno: 1996,
pagine: 653 - 663
SICI:
0002-9440(1996)149:2<653:IOHECW>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
DNA STRAND BREAKS; CELL-DEATH; FLOW-CYTOMETRY; POLY(ADP-RIBOSE) POLYMERASE; CONDENSED CHROMATIN; CLEAVAGE; FRAGMENTATION; TOLERANCE;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
36
Recensione:
Indirizzi per estratti:
Citazione:
S.J. Zunino et al., "IMMUNODETECTION OF HISTONE EPITOPES CORRELATES WITH EARLY STAGES OF APOPTOSIS IN ACTIVATED HUMAN PERIPHERAL T-LYMPHOCYTES", The American journal of pathology, 149(2), 1996, pp. 653-663

Abstract

By coupling intracellular staining with terminal deoxynucleotidyl transferase (TdT)-mediated labeling of internucleosomal DNA strand breaksin a flow cytometric assay, we observed a strong correlation between apoptosis-associated DNA strand breaks and immunoreactivity with the monoclonal antibody (MAb) B-F6 in activated human peripheral blood T lymphocytes (PBTs). Although MAb B-F6 has been reported to be specific for the cytokine interleukin-6, Western blot analysis of activated PBT lysates revealed that the predominant protein band detected by this MAb was 17 kd (p17), distinct from the 23-kd core protein and 26- to 30-kd mature glycosylated forms of interleukin-6. Immunoaffinity isolation and amino-terminal amino acid sequence analysis of p17 revealed identity with the histone H2B, a finding confirmed by Western blot analysis of purified histones and by similar staining of activated PBTs with an unrelated anti-histone MAb. Neither histone staining nor DNA strandbreakage was observed in freshly isolated PBTs; however, after T cellactivation, histone immunoreactivity appeared to precede the appearance of DNA strand breaks, with both increasing to a maximal level by day 3 after activation. Two-parameter parameter confocal immunofluorescence microscopy of histone and DNA staining confirmed a lack of histoneimmunoreactivity in viable cells and demonstrated co-localization of histone epitopes with abnormally clumped chromatin in apoptotic cells. These data indicate that alteration of histone epitope accessibility is a marker of early apoptosis and suggest that multiparameter flow cytometric analysis of intracellular epitopes may be a powerful tool in the elucidation of intracellular mechanisms of apoptosis.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/07/20 alle ore 18:42:59