METHODS
Human Subjects
It was determined using the empirical correlation between α1PI and CD4+ lymphocytes that a sample size of 2 HIV-1+ patients would be adequate two achieve a significance level with alpha = 0.05 and power of test = 0.8 between pre- and post-treatment CD4+ lymphocytes levels.
Inclusion criteria for treatment were: i) active α1PI below 11 μM; ii) one year history with CD4+ lymphocytes at levels ranging between 150 and 300 cells∕μl; iii) absence of symptoms suggestive of HIV-1 disease progression; iv) adequate suppression of virus (lymphocytes were examined for NFkB phospho-epitope staining by flow cytometry as previously described [10]. Briefly, 1x106 cells/well in 96 well plates were fixed using 1.5% paraformaldehyde, washed with PBS containing 1% BSA, and incubated at 4 °C for 10 min in 100μl ice-cold methanol. Cells were washed and incubated at 23 oC for 20 min with phosphoprotein-specific antibodies (BD Pharmingen and BD PhosFlow, San Diego, CA) directly conjugated with Alexa Fluor 647 (Molecular Probes Invitrogen, Carlsbad, CA).
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