13 TELBIVUDINE (LDT) PLUS PEG-INTERFERON (PEGIFN) IN HBEAG-POSITIVE CHRONIC HEPATITIS B – VERY POTENT ANTIVIRAL EFFICACY BUT RISK OF PERIPHERAL NEUROPATHY (PN)
2010; Elsevier BV; Volume: 52; Linguagem: Inglês
10.1016/s0168-8278(10)60015-3
ISSN1600-0641
AutoresPatrick Marcellin, Claudio Ávila, Karsten Wursthorn, Wan‐Long Chuang, G Lau, Chuan Peng, Edward Gane, Hugo Fainboim, Michael P. Manns, N.V. Naoumov,
Tópico(s)Hepatitis Viruses Studies and Epidemiology
ResumoPRESENTATIONSthus to distinguish HBV infected hepatocytes from non-infected ones.These antibodies were used to analyze the distribution and quantification of pMHC on HBV infected cells and to test their ability to deliver compounds to infected cells.Methods: Antibody secreting B cell hybridomas were generated from mice immunized with HBVc18-27/A201 and HBVe183-91/A201 pMHC monomers.The number of pMHC complexes on individual cells was enumerated by flow cytometry based quantification technique, while HBV antigen was quantified by Western blot and Quantity One Software.Confocal microscopy was used to detect intracellular delivery of fluorescent dye.Results: Culture supernatants from numerous hybridomas were screened for specificity.Five of them were able to produce antibodies that recognized T2 cells pulsed with core 18-27 (2 clones) or env 183-91 peptides (3 clones) but not T2 cells pulsed with irrelevant HLA-A0201 binding peptide.The two TCRlike antibodies could detect pMHC complexes on the surface of HepG2 cells with sensitivity lower than that of T cells (1nM versus 1pM) but they recognized the pMHC expressed naturally after endogenous processing on HBV infected cells.The binding of the antibodies was not inhibited by the presence of serum HBV antigens.The TCR-like antibodies were used to quantify pMHC complexes expressed on different cell types.On average, 1×10 5 core proteins produced one pMHC on B cells, while in hepatocytes, a single pMHC requires endogenous synthesis of approximately 1.5×10 6 proteins, directly demonstrating the limited antigen processing ability of hepatocytes.Furthermore, TCR-like antibodies could deliver the fluorescent dye to the endosomal compartment of HBV-infected hepatocytes.Conclusions: We have produced TCR-like antibodies recognizing HBV infected cells.These antibodies open the possibilities to obtain precise quantitative analysis of the peptide/HLA-class I complexes on HBV infected hepatocytes and provide the base for new therapeutic strategies to deliver drugs specifically to HBV infected cells.
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