ABSTRACT
PURPOSE: IRF-5 is a direct transducer of virus-mediated and TLR-mediated signaling pathways for the expression of cytokines and chemokines which form homodimers or heterodimers with IRF-7. However, direct IRF-5-specific monoclonal antibodies (mAbs) are not available at present. These could be used to further evaluate the functions of IRF-5. In this study, we produced and characterized three mouse mAbs to human IRF-5. The binding of IRF-5 to nuclear import proteins was first identified using a mAb. MATERIALS AND METHODS: His-tagged human IRF-5 protein spanning amino acid residues 193- 257 was used as an antigen and three mAbs were produced. The mAbs were tested with ELISA, Western blot analysis (WB), immunofluorescent staining (IF), and immunoprecipitation (IP). In addition, the nuclear import protein which carried phosphorylated IRF-5 was identified using one of these mAbs. RESULTS: MAbs 5IRF8, 5IRF10 and 5IRF24 which reacted with the recombinant His-IRF-5(193-257) protein were produced. All mAbs bound to human IRF-5, but not to IRF-3 or IRF-7. They could be used for WB, IF, and IP studies. The binding of phosphorylated IRF-5 to karyopherin-alpha1 and -beta1 was also identified. CONCLUSION: Human IRF-5-specific mAbs are produced for studying the immunologic roles related to IRF-5. Phosphorylated IRF-5 is transported to the nucleus by binding to nuclear import proteins karyopherin-alpha1 and -beta1.
Subject(s)
Animals , Humans , Mice , Antibodies, Monoclonal , Base Sequence , Cell Line , Cross Reactions , DNA Primers/genetics , Interferon Regulatory Factors/genetics , Mice, Inbred BALB C , NIH 3T3 Cells , Protein Binding , Recombinant Proteins/genetics , alpha Karyopherins/metabolism , beta Karyopherins/metabolismABSTRACT
BACKGROUND: Pneumococcus is the most common cause of acute otitis media, community acquired pneumonia and invasive bacterial diseases in children. Ninety serotypes have been identified, and the distribution differs according to geographic area and ages. The 7 valent pneumococcal protein conjugate vaccine is used widely. To evaluate the efficacy of the vaccine, it is essential to investigate the distribution of the pneumococcal serotypes. MATERIALS AND METHODS: The serotypes and antibiotic resistance of the pneumococcus isolated from 308 patients at Shinchon Severance hospital from September of 2001 to July of 2005 were analyzed. RESULTS: The pneumococcci were isolated mostly from sputum and blood, and ear discharge in the descending order. Serotyping was possible in 265 cases, and the distribution of serotypes were 19F (16.2%), 19A (12.8%), 23F (8.7%), 6B (7.9%), and 6A (7.2%). Fifty two cases were isolated from those patients less than 16 years of age and the distribution of serotypes was 19F, 19A, 23F, 14, 6B, 6A and 4. Resistance to penicillin was 64.6% in all cases and 67.3% in children. The more common serotype showed the higher rate of penicillin resistance. Multi-drug resistance was demonstrated in 64.7%. Forty three percent of the total identified serotypes were included in the 7 valent pneumococcal conjugate vaccine. And 61.5% of the serotypes identified in children were included in the vaccine. CONCLUSION: The 7 valent vaccine may be used effetively in Korea. But, further study is needed to address serotype switching after the use of the protein conjugated vaccine, which has been reported in other countries.
Subject(s)
Child , Humans , Anti-Bacterial Agents , Drug Resistance, Microbial , Drug Resistance, Multiple , Ear , Korea , Otitis Media , Penicillin Resistance , Penicillins , Pneumonia , Serotyping , Sputum , Streptococcus pneumoniae , StreptococcusABSTRACT
BACKGROUND: Pneumococcus is the most common cause of acute otitis media, community acquired pneumonia and invasive bacterial diseases in children. Ninety serotypes have been identified, and the distribution differs according to geographic area and ages. The 7 valent pneumococcal protein conjugate vaccine is used widely. To evaluate the efficacy of the vaccine, it is essential to investigate the distribution of the pneumococcal serotypes. MATERIALS AND METHODS: The serotypes and antibiotic resistance of the pneumococcus isolated from 308 patients at Shinchon Severance hospital from September of 2001 to July of 2005 were analyzed. RESULTS: The pneumococcci were isolated mostly from sputum and blood, and ear discharge in the descending order. Serotyping was possible in 265 cases, and the distribution of serotypes were 19F (16.2%), 19A (12.8%), 23F (8.7%), 6B (7.9%), and 6A (7.2%). Fifty two cases were isolated from those patients less than 16 years of age and the distribution of serotypes was 19F, 19A, 23F, 14, 6B, 6A and 4. Resistance to penicillin was 64.6% in all cases and 67.3% in children. The more common serotype showed the higher rate of penicillin resistance. Multi-drug resistance was demonstrated in 64.7%. Forty three percent of the total identified serotypes were included in the 7 valent pneumococcal conjugate vaccine. And 61.5% of the serotypes identified in children were included in the vaccine. CONCLUSION: The 7 valent vaccine may be used effetively in Korea. But, further study is needed to address serotype switching after the use of the protein conjugated vaccine, which has been reported in other countries.