publication . Thesis . 2006

Specificity and polyreactivity of the antibody response during natural HIV-1 infection

Wang, Xin;
Open Access English
  • Published: 01 Jan 2006
  • Country: Canada
Abstract
The specificity and polyreactivity of the antibody response in natural HIV-1 infection were studied. First, to investigate the overall antibody response, overlapping linear peptides were used to screen sera taken from HIV-1-infected individuals. The polyclonal antibody response was relatively stable during long-term infection, compared with acute infection, and mostly directed against immunodominant regions. Low level, transient antibody responses were detected against membrane proximal external region of gp41. To test if these antibodies are neutralizing, an affinity purification method was developed to isolate these serum antibodies. Second, in terms of polyre...
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Introduction ...............................................................................................5..1..

Materials and methods ...............................................................................5..3. Reagents ...............................................................................................5..3... Antibody affinity purification - Trial 1 .....................................................54 Antibody affinity purification - Trial 2 ...................................................5..5 Direct ELISA for characterizing purified IgG binding to specific peptide ....................................................................................................5..6. Neutralization assay ...............................................................................5..7.

Discussion ...................................................................................................9..2 Figure 4-2: Regression of ELISA signal from 10 nM MAb CL 15 on net charge of synthetic peptides.................................................................................8..4..

Figure 4-3: Neutralization of pseudotyped HIV-I HxB2 by MAbs CL 15. 4E 10 and 15E......................................................................................................... 87 Figure 4-4: Figure 4-5: Anti-CL Abs as a marker for autoreactivity during HIV-1 infection...........88 Cardiolipin reactivity in sera (1 in 50 dilution)...........................................9.0 Table 2-1: Table of HIV- 1 MN Env peptides obtained from the NIH AIDS Research & Reference Reagent Program.....................................................30 Table 3-1: Summary of IC50 in neutralization assay with bNt serum #6. Merck serum (negative control)...........................................................................7..4.

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