ABSTRACT
<p><b>OBJECTIVE</b>To prepare highly specific chicken egg yolk IgY antibody against human papillomavirus 16 type L1 main capsid protein (HPV16L1) for detection of HPV16L1.</p><p><b>METHODS</b>Purified HPV16L1 protein was used to immunize the hens, from which the eggs were collected since one week after the first immunization. The egg yolk was separated and the IgY antibody purified by PEG-6000 method. The bioactivity of the antibody was tested using enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry was performed to detect the HPV16L1 in the CHO cells transfected with the recombinant pcDNA-EGFP-HPV16L1 plasmid (containing EGFP-HPV16L1 fusion gene) for assessing the specific affinity of IgY to HPV16L1.</p><p><b>RESULTS</b>After 3 immunizations of the hens, the titer of the purified IgY antibody against HPV16L1 from the egg yolk reached 1:10240. The IgY bound specifically to the EGFP-HPV16L1 protein expressed in the transfected CHO cells.</p><p><b>CONCLUSION</b>High titer IgY can be prepared by immunization of the hens with HPV16L1 protein, and the prepared IgY can be used for HPV16L1 detection at the cellular level.</p>
Subject(s)
Animals , Cricetinae , Humans , Mice , Antibodies, Viral , Allergy and Immunology , Antibody Specificity , Allergy and Immunology , CHO Cells , Capsid Proteins , Genetics , Allergy and Immunology , Metabolism , Chickens , Cricetulus , Enzyme-Linked Immunosorbent Assay , Green Fluorescent Proteins , Genetics , Allergy and Immunology , Metabolism , Immunization , Methods , Immunoglobulins , Allergy and Immunology , Immunohistochemistry , Mice, Inbred C57BL , Oncogene Proteins, Viral , Genetics , Allergy and Immunology , Metabolism , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Metabolism , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To investigate the relationship between HER-2 expression and the efficacy of neoadjuvant chemotherapy in local advanced breast cancer.</p><p><b>METHODS</b>Different neoadjuvant chemotherapy regimens, namely CMF, CEF, and NEF, were administered in 132 patients with local advanced breast cancer for 2 cycles, each lasting for 28 days. According to the criteria recommended by WHO, the efficacy and safety of the regimens were evaluated after two cycles of neoadjuvant chemotherapy. HER-2 expression was examined by immunohistochemistry using specific monoclonal antibodies before chemotherapy and after surgery.</p><p><b>RESULTS</b>The overall response rate (RR) of CMF, CEF, and NEF regimens were 39.5% (17/43), 54.3% (25/46) and 72.1% (31/43), with incidence of leukopenia of 34.9% (15/43), 58.7% (27/46) and 60.5% (26/43), respectively. Other adverse effects including decreased hemoglobin (Hb) level, thrombocytopenia, gastrointestinal irritation and alopecia were similar between the 3 groups (P>0.05). No significant variation in HER-2 expression occurred after administration of the 3 regimens. The overall RR to CMF regimen in HER-2-negative breast cancer patients was significantly higher than that in HER-2-positive patients, but showed no significant difference with CEF and NEF regimens.</p><p><b>CONCLUSION</b>HER-2 expression is not decreased after neoadjuvant chemotherapy in breast cancer patients, and HER-2-positive breast cancer can be resistant to CMF regimen, but not to CEF and NEF regimens.</p>