Characterization of a Monoclonal Antibody, HTA28, Recognizing a Histone H3 Phosphorylation Site as a Useful Marker of M-phase Cells

ArticleinJournal of Histochemistry and Cytochemistry 52(11):1503-9 · December 2004with25 Reads
DOI: 10.1369/jhc.4A6285.2004 · Source: PubMed
Mitosis is a valuable indicator of active tissue proliferation but, other than morphological characteristics, there have hitherto been no markers available to detect only M-phase cells. However, a newly established monoclonal antibody (MAb), HTA28, recognizing histone H3 (H3) harboring phosphoserine 28, allows visualization with mitotic chromosomal condensation. In this study we investigated the use of HTA28 for immunohistochemical (IHC) detection of M-phase cells in the regenerating rat liver after partial hepatectomy (PH). Groups of three to five rats were sacrificed at intervals up to 72 hr after PH and proliferation was then assessed by IHC staining using HTA28 and other markers. The temporal pattern of the HTA28 staining index (SI) was very similar to that for the mitotic index (MI), also showing similarities to the bromodeoxyuridine (BrdU) labeling index (LI) with a time lag. The HTA28 SI proved to be higher than MI at every time point in line with HTA28 immunoreactivity maintained for all stages of M-phase. The spatial distribution of HTA28-positive cells corresponded with those of other proliferative cell markers. These therefore provide strong evidence for the applicability of HTA28 as an M-phase marker. We also showed that antigenicity for HTA28 is lost if tissue is not immediately fixed after sampling.
    • "Erkan et al. [39] compared Ki-67-positive proliferative index in IM and chronic gastritis and showed a significantly higher index in IM. Histone H3 is phosphorylated at serine 28 during the M phase in the cell cycle, being detected with HTA28 antibody [24] . In G-type glands harboring no IM, H3S28ph-positive cells were not increased with H. pylori infection , which was unexpectedly not altered by eradication in contrast to the proliferative zone characterized as the Ki-67-positive region. "
    Full-text · Article · Jan 2016
    • "Histone H3 is phosphorylated at position S28 during M phase [9, 10]. An available mAb binds to only the phosphorylated form of histone H3 at this position, making it a good marker to estimate the time to enter M phase. "
    [Show abstract] [Hide abstract] ABSTRACT: Peripheral T cells are in G0 phase and do not proliferate. When they encounter an antigen, they enter the cell cycle and proliferate in order to initiate an active immune response. Here, we have determined the first two cell cycle times of a leading population of CD4(+) T cells stimulated by PMA plus ionomycin in vitro. The first cell cycle began around 10 h after stimulation and took approximately 16 h. Surprisingly, the second cell cycle was extremely rapid and required only 6 h. T cells might have a unique regulatory mechanism to compensate for the shortage of the gap phases in cell cycle progression. This unique feature might be a basis for a quick immune response against pathogens, as it maximizes the rate of proliferation.
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    • "Liver slices were fixed in 10% neutral buffered formalin and were processed in a routine manner. Immunohistochemical staining for GST-P (MBL Co. Ltd., Nagoya, Japan ) and Ki-67 (Clone MIB-5, DakoCytomation, Glostrup, Denmark) and analysis of GST-P-positive foci were performed as described previously (Hirata et al., 2004; Sakai et al., 2002). To investigate the difference in cell proliferation activity between GST-P-positive foci and the surrounding tissues, serially sectioned slides were stained for GST-P and Ki-67. "
    [Show abstract] [Hide abstract] ABSTRACT: We aimed to investigate the combined effect of organochlorine pesticides heptachlor (HEP) and hexachlorobenzene (HCB) by using a medium-term rat liver bioassay. Male F344 rats were initially administered diethylnitrosamine (DEN, 200mg/kgi.p.); after a 2-week non-dosing period, they were given diets containing HEP (5 or 25ppm), HCB (70 or 350ppm), or their mixtures (5 and 70ppm or 25 and 350ppm) for 6weeks. All rats were subjected to partial hepatectomy at week 3 and killed at week 8. We observed additive or synergistic effects of HEP and HCB in groups treated with mixtures of these pesticides. Number and area of preneoplastic foci positive for glutathione S-transferase placental form (GST-P) were consistently higher in these groups than the sum of individual values in the groups treated with HEP or HCB alone. Consistent with these findings, HEP and HCB had additive or synergistic effects on cell proliferation induction within the preneoplastic foci and cytochrome P450 (CYP) 2B1 and 3A1 induction, which may lead to more efficient metabolic activation of HEP and HCB. On the basis of these findings, we conclude that HEP and HCB have additive and synergistic effects on the development of GST-P-positive foci and that higher risks are associated with a combination of residual organochlorine pesticides in foods than with individual residual organochlorine pesticides.
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