Formulation of Poly(DL-Lactide-Co-Glycolide) Microspheres and Their Ingestion by Bovine Leukocytes

¹ Department of Animal Sciences, University of Maryland, College Park 20742
² Immunology and Disease Resistance Laboratory, USDA, Agricultural Research Service, Beltsville, MD 20705

This study determined optimal parameters for producing controlled-release microspheres and examined their suitability for vaccines via ingestion by bovine leukocytes. Microspheres elicit an immune response when ingested by antigen-presenting cells and provide sustained exposure of antigen to sensitized cells. Ingestion of microspheres is determined by their size (<10 µm), and antigen release is governed by composition. Poly(DL-lactide-co-glycolide) microspheres were prepared using different polymer concentrations, stir rates, emulsifier concentrations, and emulsifier molecular masses. Microspheres that were <10 µm were prepared using a 6% 50:50 lactide to glycolide polymer solution emulsified at 12,000 rpm in a low molecular mass, 5% polyvinyl alcohol solution. Microspheres that were >20 µm were prepared using a 10% 85:15 lactide to glycolide polymer solution emulsified at 1200 rpm in a low molecular mass, 3% polyvinyl alcohol solution. Small microspheres released 90% of the antigen after 7 d, and large microspheres released only 24% of the antigen after 56 d. Both monocytes and neutrophils selectively ingested small microspheres that were opsonized with normal bovine serum (heated and unheated). Ingestion of microspheres that had been opsonized with fetal bovine serum (heated and unheated) was minimal. Lymphocytes did not ingest microspheres. Ingestion of small microspheres by bovine monocytes and sustained release of antigen by large microspheres suggested that microspheres have the ability to produce a sustained immune response with a single injection.

Key Words: microsphere • formulation • ingestion • leukocytes

Submitted on November 21, 1995
Accepted on April 1, 1996