The use of monoclonal anti-Thy1 IgG1 for the targeting of liposomes to AKR-A cells in vitro and in vivo.
A number of SH-containing proteins or protein derivatives were coupled to small unilamellar liposomes. These were composed of distearoylphosphatidylcholine (DSPC), dipalmitoylphosphatidylethanolamine (DPPE) and cholesterol (1:1, phospholipid/cholesterol molar ratio) and activated (DPPE moiety) with the heterobifunctional reagents N-hydroxysuccinimide ester of iodoacetic acid (hydroxysuccinimide iodoacetate), N-succinimidyl-4-(2-bromoacetylamino)benzoate (SBAB) or N-succinimidyl-3-(2-pyridyldithio)proprionate (SPDP). DPPE was activated with the reagents before or after its incorporation into liposomes. Protein coupling values varied widely depending on the reagent and the protein used, but were highest in the case of SPDP-activated liposomes and SPDP-modified immunoglobulin G (IgG). Monoclonal anti-Thy1 125I-IgG1-bearing liposomes (SPDP- or SBAB-activated) containing quenched carboxyfluorescein were incubated under a variety of conditions with mouse AKR-A cells expressing the cross-reactive Thy 1.1 antigen. The following observations were made; (a) binding of intact liposomes to the cells at 4 degrees C reached plateau values after about 1 h with at least 70% of the liposomes used being capable of associating with the target cells; (b) binding of liposomes to AKR-A cells was much more pronounced than when using another cell line (EL4-Tc); (c) binding to AKR-A cells could be effected with as little as 1.3 molecules (average) of IgG1 per vesicle; (d) binding was inhibited only modestly by the presence of 50% mouse plasma; (e) stability of IgG1-bearing liposomes in terms of entrapped solute and IgG1 retention in the presence of plasma at 37 degrees C was maintained quantitatively for at least 5.5 h, and by 24 h, 54% of the IgG1 was still associated with the liposomes. AKR mice were injected intravenously with 99mTc-labelled AKR-A cells and 2.5 min later with anti-Thy1 125I-IgG1-bearing liposomes containing quenched carboxyfluorescein and 111In-Ca-DTPA or with similar liposomes devoid of IgG1. In parallel experiments, AKR mice received either of the liposome preparations without previous injection of cells. On the basis of patterns of quenched carboxyfluorescein, 111In and 125I-clearance from the circulation, of 99mTc levels in the blood and of values of 111In in the liver and spleen, it appeared that IgG1-bearing liposomes were capable of binding to their target cells in the vasculature. Such binding accelerated the clearance of interacting moieties (i.e., AKR-A cells and liposomes). The present results suggest that targeting of liposomes to circulating in vivo is feasible.
D000818:Animals; D001769:Blood; D002460:Cell Line; D003080:Cold Temperature; D003429:Cross Reactions; D004304:Dosage Forms; D007074:Immunoglobulin G; D007518:Isoantibodies; D008081:Liposomes; D051379:Mice; D013997:Time Factors
D004304:Dosage Forms; D007074:Immunoglobulin G; D007518:Isoantibodies; D008081:Liposomes; C035678:anti-Thy antibody