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A new proliferation-associated nuclear antigen detectable in paraffin-embedded tissues by the monoclonal antibody
H Kreipe1, H J Heidebrecht, S Hansen
1Institute of Pathology and Hematopathology, University of Kiel, Germany.
Insights
A new monoclonal antibody, Ki-S1, identifies proliferating cells by targeting a nuclear antigen. This antibody is effective in various tissue preparations and shows promise for studying cell growth in normal and diseased states.
Area of Science:
- Immunology
- Cell Biology
- Pathology
Background:
- Cell proliferation is a fundamental biological process.
- Accurate detection of proliferating cells is crucial for understanding normal development and disease progression, including cancer.
- Existing markers for cell proliferation have limitations in their application and specificity.
Purpose of the Study:
- To develop and characterize a novel monoclonal antibody (Ki-S1) for detecting proliferating cells.
- To evaluate the utility of Ki-S1 in immunohistochemistry and flow cytometry.
- To assess the antigen's properties and potential as a proliferation marker.
Main Methods:
- Generation of a monoclonal antibody (Ki-S1).
- Immunohistochemical staining on formalin-fixed and frozen tissues.
- Flow cytometric analysis of DNA content and antigen expression in lymphocytes.
- Western blot analysis to determine antigen molecular mass.
Main Results:
- Ki-S1 antibody specifically recognizes a nuclear antigen in proliferating cells.
- The antigen is detectable in both frozen and routinely processed tissues.
- Peripheral blood lymphocytes show increased antigen expression upon mitogen stimulation.
- Flow cytometry confirmed antigen expression in G1 to G2/M phases, but not G0.
- A distinct cytoplasmic reactivity was observed in pancreatic Langerhans islands.
- The nuclear antigen has a molecular mass of 160 kDa, differentiating it from other markers.
Conclusions:
- Monoclonal antibody Ki-S1 is a valuable tool for identifying proliferating cells.
- Its resistance to formalin fixation and applicability to various tissue types enhance its utility.
- Ki-S1 offers a potentially novel marker for in situ studies of cell proliferation in diverse biological contexts.
Abstract:
A monoclonal antibody (Ki-S1) has been raised that reacts with the nuclei of proliferating cells. The antigen recognized is resistant to formalin fixation and can be detected in frozen tissues as well as in routinely processed specimens. In immunohistochemistry, nuclear staining can be seen in those tissues and cellular compartments known to be actively proliferating. Peripheral blood lymphocytes are negative but show a strong increase in antigen expression after mitogen stimulation. Flow cytometric determination of DNA content and antigen expression revealed negativity of G0 cells and positivity of G1 to G2/M cells. A cytoplasmic co-reactivity, not associated with proliferation, was confined to Langerhans islands of the pancreas. The nuclear localized antigen has a molecular mass of 160 kd and therefore seems to be different from all other known immunohistochemical markers of proliferating cells. We conclude that the monoclonal antibody Ki-S1 might provide a useful tool for studying cell proliferation in situ under normal and pathological circumstances.
