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1.
Chinese Journal of Virology ; (6): 383-389, 2008.
Article in Chinese | WPRIM | ID: wpr-334791

ABSTRACT

Human adenovirus serotype 5 (HAd5) infect dendritic cells with low efficiency which restricts the use of HAd5 as an antigen carrying vector in such cells. Aiming to find a novel strategy to detour the traditional method for more convenient clinical use, peripheral blood mononuclear cells isolated from Chinese rhesus macaques were chosen as the target cells for HAd5. In vitro infection protocol was optimized which indicated centrifugation at 1000g could ease the entry of adenovirus. By this protocol, CD14 positive monocytes were infected at high efficiencies (> 80%), and about 10% of natural killer cells were infected; while T and B lymphocytes were rarely infected. Interestingly and importantly, it was the first time to report that in our in vitro study monocytes were more susceptible to HAd5-EGFP in macaques with higher preexisting vector specific neutralizing antibody titers. This phenomenon indicates an expansion of application of adenovirus based vectors for vaccine development and clinical use, especially for the population with preexisting neutralizing antibodies.


Subject(s)
Animals , Female , Humans , Male , Adenoviridae , Classification , Genetics , Allergy and Immunology , Antibodies, Viral , Blood , Genetic Vectors , Lipopolysaccharide Receptors , Macaca mulatta , Monocytes , Virology , Neutralization Tests , Recombination, Genetic , Serotyping
2.
Journal of Southern Medical University ; (12): 425-427, 2006.
Article in Chinese | WPRIM | ID: wpr-255294

ABSTRACT

<p><b>OBJECTIVE</b>To prepare and identify monoclonal antibodies (mAbs) against Helicobacter pylori (Hp).</p><p><b>METHODS</b>BALB/c mice were immunized with the supernatant and precipitation of cultured Hp after ultrasonication and mAbs were obtained by means of hybridoma technique. The resultant mAbs was evaluated for subtype, titer, affinity, and further identified with Lpp20, HspA, urease A, CagA, urease B, and catalase prepared by recombinant expression.</p><p><b>RESULTS</b>Totally 34 hybridoma cell lines were established which secreted specific mAbs, including 31 against the supernatant and 3 against the precipitation of Hp, and the prepared mAbs showed specific reaction against Lpp20 (3 strains), HspA (2 strains), urease A (4 strains), CagA (1 strain), urease B (5 strains), and catalase (2 strains) antigens, respectively. The mAbs was all identified as immunoglobulin G1 (IgG1) and theirs titer in the culture supernatant and ascites was 1:16 to 1:32 and 1:32000 to 1:64000 respectively with affinity constants (K(aff)) ranging from 1 x 10(-10) to 5.2 x 10(-12) mol/L.</p><p><b>CONCLUSION</b>The mAbs specially against Hp have been obtained, which may facilitate further study of detection and vaccine development of Hp.</p>


Subject(s)
Animals , Female , Humans , Mice , Antibodies, Bacterial , Allergy and Immunology , Antibodies, Monoclonal , Allergy and Immunology , Antibody Specificity , Helicobacter pylori , Allergy and Immunology , Hybridomas , Allergy and Immunology , Mice, Inbred BALB C
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