Pharmaceutical Research

, Volume 29, Issue 9, pp 2565–2577

Optimization of Stability, Encapsulation, Release, and Cross-Priming of Tumor Antigen-Containing PLGA Nanoparticles

  • Shashi Prasad
  • Virginia Cody
  • Jennifer K. Saucier-Sawyer
  • Tarek R. Fadel
  • Richard L. Edelson
  • Martin A. Birchall
  • Douglas J. Hanlon
Research Paper

Abstract

Purpose

In order to investigate Poly (lactic-co-glycolic acid) (PLGA) nanoparticles (NP) as potential vehicles for efficient tumor antigen (TA) delivery to dendritic cells (DC), this study aimed to optimize encapsulation/release kinetics before determining immunogenicity of antigen-containing NP.

Methods

Various techniques were used to liberate TA from cell lines. Single (gp100) and multiple (B16-tumor lysate containing gp100) antigens were encapsulated within differing molecular weight PLGA co-polymers. Differences in morphology, encapsulation/release and biologic potency were studied. Findings were adopted to encapsulate fresh tumor lysate from patients with advanced tumors and compare stimulation of tumor infiltrating lymphocytes (TIL) against that achieved by soluble lysate.

Results

Four cycles of freeze-thaw + 15 s sonication resulted in antigen-rich lysates without the need for toxic detergents or protease inhibitors. The 80KDa polymer resulted in maximal release of payload and favorable production of immunostimulatory IL-2 and IFN-γ. NP-mediated antigen delivery led to increased IFN-γ and decreased immunoinhibitory IL-10 synthesis when compared to soluble lysate.

Conclusions

Four cycles of freeze-thaw followed by 15 s sonication is the ideal technique to obtain complex TA for encapsulation. The 80KDa polymer has the most promising combination of release kinetics and biologic potency. Encapsulated antigens are immunogenic and evoke favorable TIL-mediated anti-tumor responses.

Key Words

antigen delivery dendritic cell immunotherapy molecular weight nanoparticles 

Abbreviations

Ags

antigens

APC

antigen presenting cell

ATCC

American Type Culture Collection

BCA

bicinchonic acid

BMDC

bone marrow dendritic cells

BSA

bovine serum albumin

CBA

cytometric bead array

CD

cluster of differentiation

CTL

cytotoxic T cells

DC

dendritic cell

FT + S

freeze-thaw + sonication

GM-CSF

granulocyte macrophage-colony stimulating factor

HNSCC

head and neck squamous cell carcinoma

IFN-γ

interferon-γ

IL

interleukin

LPS

lipopolysaccharide

MP

microparticles

MW

molecular weight

NP

nanoparticles

PBS

phosphate buffered saline

PLGA

poly (lactic-co-glycolic acid)

PVA

polyvinyl alcohol

RT

room temperature

SDS-PAGE

sodium dodecyl sulfate polyacrylamide gel electrophoresis

SEM

scanning electron microscopy

SPSS

Statistical Package for the Social Sciences

TA

tumor associated antigen

Th

T-helper

TIL

tumor infiltrating lymphocytes

TNF-α

tumor necrosis factor-α

WB

Western blot

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Shashi Prasad
    • 1
  • Virginia Cody
    • 1
  • Jennifer K. Saucier-Sawyer
    • 2
  • Tarek R. Fadel
    • 2
  • Richard L. Edelson
    • 1
  • Martin A. Birchall
    • 3
  • Douglas J. Hanlon
    • 1
    • 4
  1. 1.Department of DermatologyYale UniversityNew HavenUSA
  2. 2.Department of Biomedical EngineeringYale UniversityNew HavenUSA
  3. 3.UCL Center for Stem Cells & Regenerative Medicine & UCL Ear InstituteRoyal National Throat Nose & Ear HospitalLondonUK
  4. 4.Yale UniversityNew HavenUSA

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