ABSTRACT
<p><b>BACKGROUND</b>Infections caused by gram-negative bacteria (GNB) often lead to high mortality in common clinical settings. The effect of traditional antibiotic therapy is hindered by drug-resistant bacteria and unneutralizable endotoxin. Few effective methods can protect high risk patients from bacterial infection. This study explored the protection of adeno-associated virus 2 (AAV2)-bacteriacidal permeability increasing protein 700 (BPI(700))-fragment crystallizable gamma one 700 (Fc gamma1(700)) chimeric gene transferred mice against the minimal lethal dose (MLD) of E. coli and application of gene therapy for bacterial infection.</p><p><b>METHODS</b>After AAV2-BPI(700)-Fc gamma1(700) virus transfection, dot blotting and Western blotting were used to detect the target gene products in Chinese hamster ovary-K1 cells (CHO-K1cells). Reverse transcription-polymerase chain reaction and immunohistochemical assay were carried out to show the target gene expression in mice. Modified BPI-enzyme linked immunosorbent assay was used to identify the target gene products in murine serum. The protection of BPI(700)-Fc gamma1(700) gene transferred mice was examined by survival rate after MLD E. coli challenge. Colony forming unit (CFU) count, limulus amebocyte lysate kit and cytokine kit were used to quantify the bacteria, the level of endotoxin, and proinflammatory cytokine.</p><p><b>RESULTS</b>BPI(1-199)-Fc gamma1 protein was identified in the CHO-K1 cell culture supernatant, injected muscles and serum of the gene transferred mice. After MLD E. coli challenge, the survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice (36.7%) was significantly higher than that of AAV2-enhanced green fluorescent protein (AAV2-EGFP) gene transferred mice (3.3%) and PBS control mice (5.6%). The survival rate of AAV2-BPI(700)-Fc gamma1(700) gene transferred mice treated with cefuroxime sodium was 65.0%. The bacterium number in main viscera, the levels of endotoxin and proinflammatory cytokine (tumor necrosis factor-alpha and interleukin-1beta) in serum of the AAV2-BPI(700)-Fc gamma1(700) gene transferred mice were markedly lower than that of PBS control mice (P < 0.01).</p><p><b>CONCLUSIONS</b>AAV2-BPI(700)-Fc gamma1(700) gene transferred mice can resist MLD E. coli infection through expressing BPI(1-199)-Fc gamma1 protein. Our findings suggested that AAV2 mediated BPI(700)-Fc gamma1(700) gene delivery could be used for protection and treatment of clinical GNB infection in high-risk individuals.</p>
Subject(s)
Animals , Cricetinae , Mice , Anti-Bacterial Agents , Therapeutic Uses , Antimicrobial Cationic Peptides , Blood Proteins , CHO Cells , Dependovirus , Genetics , Disease Models, Animal , Escherichia coli Infections , Therapeutics , Gene Transfer, Horizontal , Genetic Therapy , Mice, Inbred BALB C , Proteins , Genetics , Receptors, IgG , Genetics , Recombinant Fusion Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To identify the expression and distribution of macrophage colony stimulating factor (M-CSF) in brains of PDAPPV717I transgenic mice.</p><p><b>METHODS</b>We detected the expression and distribution of M-CSF mRNA in brains of PDAPPV717I transgenic mice by using hybridization in situ and immunohistochemical staining.</p><p><b>RESULTS</b>Expression of M-CSF mRNA was significantly higher in brains of PDAPPV717I transgenic mice than that in non-transgenic mice, and M-CSF mRNA in brain was mainly produced by reactive astrocytes.</p><p><b>CONCLUSION</b>The results indicate that astrocytes play an important role in the onset/development of neuropathology of Alzheimer's disease.</p>
Subject(s)
Animals , Female , Male , Mice , Alzheimer Disease , Genetics , Metabolism , Brain , Metabolism , Immunohistochemistry , In Situ Hybridization , Macrophage Colony-Stimulating Factor , Genetics , Mice, Transgenic , RNA, Messenger , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the availability of serum level of macrophage clony stimulating factor (M-CSF) as a marker for early diagnosis of Alzheimer's disease (AD).</p><p><b>METHODS</b>The serum levels of M-CSF in 70 patients with AD, 52 healthy controls, 22 patients with VAD (vascular dementia) were measured and the serum levels of IL-1 beta, IL-6, TNF-alpha in 32 patients with AD and 20 controls were measured as well.</p><p><b>RESULTS</b>Serum levels of M-CSF were significantly elevated in patients with AD when compared with healthy controls (P < 0.01) and VAD controls (P < 0.05) respectively. At the early stage of mild dementia and middle dementia, serum levels of M-CSF were significantly elevated, but at the later stage of severe dementia, they returned to normal level. Serum levels of IL-1 beta were significantly elevated in AD patients compared with controls (P < 0.05), and serum levels of TNF-alpha and IL-6 were within the normal range in patients with AD.</p><p><b>CONCLUSIONS</b>The results suggest that serum M-CSF level may provide a convenient and sensitive means for the early diagnosis of Alzheimer's disease.</p>