ABSTRACT
<p><b>OBJECTIVE</b>To prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovirus 71.</p><p><b>METHODS</b>Two peptides, SP55 and SP70, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to establish the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test.</p><p><b>RESULTS</b>Two high titered anti-VP1 antibodies secreted by the hybridoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1:16 respectively.</p><p><b>CONCLUSION</b>Two high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.</p>
Subject(s)
Animals , Humans , Male , Mice , Antibodies, Monoclonal , Allergy and Immunology , Antibodies, Viral , Allergy and Immunology , Capsid Proteins , Allergy and Immunology , Enterovirus , Chemistry , Allergy and Immunology , Enterovirus Infections , Diagnosis , Allergy and Immunology , Virology , Enzyme-Linked Immunosorbent Assay , Mice, Inbred BALB C , Neutralization TestsABSTRACT
<p><b>OBJECTIVE</b>To study the effect of SiO(2) on the expression of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) in human alveolar macrophages (AMs) associated with the pathogenesis of silicotic fibrosis.</p><p><b>METHODS</b>AMs were collected from a silicotic patient by bronchoalveolar lavage, and exposed to SiO(2) (50 microg/ml), and cultured in DMEM without serum for different time (2, 6, 12, 18, 24, 36 h). Immunocytochemical method was used to detect the level of expression of MMP-9 and TIMP-1 in AMs.</p><p><b>RESULTS</b>The expression of MMP-9 in AMs exposed to silica was up-regulated, and reached the peak at 18 h [average optical density: (0.440 +/- 0.021) vs (0.390 +/- 0.011), P < 0.05]. After that, the expression reduced markedly. However, the expression of TIMP-1 of AMs were not significantly different from the control group [average optical density: (0.175 +/- 0.019) vs (0.162 +/- 0.044), P > 0.05].</p><p><b>CONCLUSION</b>SiO(2) could induce up-expression of MMP-9 in AMs. Degradation of basement membrane by MMP-9 produced by AMs at early stage of lung injury may associate with the immigration of various cells including lung fibroblasts into the injured region.</p>