We tested an affinity hemodialysis technique designed to efficiently remove HIV and toxic viral proteins from blood. Miniature polyethersulfone hollow-fiber dialysis cartridges (200-500 nm pore) were packed with anti-HIV antibodies covalently coupled to agarose beads and sealed inside the cartridge. Cell culture fluids, plasma, or infected blood (7-15 ml) containing HIV-1 were circulated over the cartridge at 0.7-10 ml/min and the rate of removal of HIV measured by PCR and p24 ELISA. The technique removed up to 98% of HIV-1 particles from cell culture supernatants. Affinity hemodialysis also efficiently cap-tured cultured HIV from human blood plasma (90%) and native HIV from infected blood (83% to 100%). Viral capture followed first-order kinetics (t 1/2 ס 2.8 h). Variations in antibody type, matrix linkage (protein G versus direct coupling), bead pore size, and temperature of operation (25-37°C) had only small effects. Although some binding was nonspecific, direct binding to the immobilized antibodies appeared to be the predominant mechanism.
Background: HIV-1 gp120 may play a role in the progression from HIV infection to AIDS. Aims: We investigated affinity hemodialysis for removing gp120 from cell culture and whole blood. Methods: Anti-gp120 antibodies covalently coupled to agarose beads were packed into columns or hollow-fiber hemodialysis cartridges. Supernatants from HIV-infected HL2/3 cells or gp120 containing whole blood were pumped over the columns and gp120 measured by ELISA. Results: Anti-gp120 agarose removed ∼90% of HIV-1 gp120 from HL2/3 cultures in 30–60 min. Capture was antibody-dependent (F105 > IDX 1121 > ABI 13-108). Affinity hemodialysis also efficiently captured gp120 from buffer in a first-order, flow-rate-dependent fashion (t½ = 13 min at 0.9 ml/min). Clearance was faster than calculated diffusion (t½ ∼2.5 h) suggesting significant convective transport. gp120 removal from blood was slower (t½ = 1.4 h). Conclusion: Affinity hemodialysis efficiently clears gp120 from cell culture fluids and blood and may be useful in slowing the progression to AIDS.
Fifteen long-term hemodialysis patients suffering from stable anemia received recombinant human erythropoietin (r-huEPO). The hormone was given intravenously at the end of each dialysis session starting with a dose of 24 IU/kg. This dose was doubled when hemoglobin levels did not rise within 2 weeks. The number of reticulocytes started to increase after 14 days of treatment. The hematocrit rose from baseline values of 23.7 +/- 1.2% to 32.4 +/- 1.3% after 24 weeks of treatment. In parallel, hemoglobin values increased from 7.3 +/- 0.3 g/100 ml to 10.1 +/- 0.4 g/100 ml. As for side effects, 3 patients developed hypertension and 2 patients suffered from occlusions of their arterio-venous fistulas. There was no evidence of major organ dysfunctions, toxic effects, allergic reactions, or antibody formation. These data show that r-HuEPO is able to correct the anemia of patients undergoing hemodialysis treatment.
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