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. 2008;1:40-45.
doi: 10.2174/1875039700801010040.

Quantitative Proteomic Analysis of Formalin Fixed Paraffin Embedded Oral HPV Lesions From HIV Patients

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Free PMC article

Quantitative Proteomic Analysis of Formalin Fixed Paraffin Embedded Oral HPV Lesions From HIV Patients

Mohit Raja Jain et al. Open Proteomics J. .
Free PMC article

Abstract

Human immunodeficiency virus (HIV) infection is associated with dysplastic changes in oral human papilloma virus (HPV) lesions, suggesting changes in keratinocytes. In the present study, we seek to identify proteomic changes in oral HPV lesions between HIV(+) and HIV(-) patients. While fresh tissues represent the most desirable samples for proteomic investigations, they are often difficult to obtain in large numbers under clinical settings. We therefore have developed a new method to identify protein changes in formalin fixed and paraffin-embedded (FFPE) oral HPV lesions utilizing iTRAQ™ technology in conjunction with Liquid Tissue® sample preparation method. Using this method, we identified nine proteins that were differentially expressed in oral HPV lesions as a result of HIV infection. The quantitative proteomic method presented here will be valuable for others who plan to analyze FFPE tissues.

Figures

Fig. (1)
Fig. (1)
Examples of quantification and identification of proteins. Cytokeratin 17 (A-D) was downregulated and histone H2A.z (E and F) was upregulated in HPV(+) HIV(+) samples. Two peptides corresponding to cytokeratin 17 (A and C) are shown to indicate similar trends in peptide quantification. Peptide sequences were deduced from the MS/MS spectra, (B, D and F) based on the observation of continuous series of either N-terminal (b series) or C-terminal (y series) ions. Quantification of peptides was based on the relative iTRAQ reporter peak areas (A, C and E).
Fig. (2)
Fig. (2)
Immunoperoxidase staining for cytokeratin 17. Variable staining of suprabasal keratinocytes is shown in papillomas removed from the oral mucosa. The overall observation is that of decreased staining intensity of oral papilloma keratinocytes in the HIV(+) patients (B) as compared to papillomas found in the HIV(-) patients (A) (100X), corroborating the iTRAQ quantification trend.

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