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The identification of amyloid P-component (protein AP) in normal cultured human fibroblasts
Abstract:
The amyloid protein, AP, has not previously been detected in normal or nonamyloidotic tissues. In this study, human fibroblasts were examined for the presence of AP by immunofluorescence technique. Normal human skin fibroblasts were cultured in Eagle's minimum essential medium for varying periods of time. Antiserum was prepared in rabbits using AP isolated from amyloid-rich tissues. Using indirect immunofluorescence and anti-AP, fluorescent material was found to be distributed in a punctate pattern throughout the cytoplasm with sparing of the nucleus. The fluorescent reaction was speicifically inhibited by absorption of anti-AP with AP-positive serum but not by addition of AP-negative human cord serum. Thus, AP may have a fibroblast origin and a relationship to normal connective tissue.
Insights
Amyloid protein (AP) was detected in normal human fibroblasts using immunofluorescence. This suggests AP may originate from fibroblasts and relate to connective tissue.
Area of Science:
- Biochemistry
- Cell Biology
- Immunology
Background:
- Amyloid protein (AP) has not been previously identified in normal or nonamyloidotic tissues.
- The cellular origin and normal function of AP remain largely uncharacterized.
Purpose of the Study:
- To investigate the presence and localization of AP in normal human fibroblasts.
- To determine if AP has a fibroblast origin and a potential role in connective tissue.
Main Methods:
- Culturing of normal human skin fibroblasts.
- Preparation of rabbit antiserum against purified AP.
- Indirect immunofluorescence technique using anti-AP antiserum.
Main Results:
- AP was detected within the cytoplasm of human fibroblasts in a punctate pattern.
- The specific binding of anti-AP was confirmed by absorption experiments.
- Fluorescent staining was specifically inhibited by AP-positive serum.
Conclusions:
- Amyloid protein (AP) can be detected in normal human fibroblasts.
- Findings suggest a potential fibroblast origin for AP.
- AP may be associated with normal connective tissue functions.