Liposome‐bound APO2L/TRAIL is an effective treatment in a rabbit model of rheumatoid arthritis

Abstract

Objective. We previously observed that T lymphocytes present in synovial fluid (SF) from patients with rheumatoid arthritis (RA) were sensitive to APO2L/ TRAIL. In addition, there was a drastic decrease in the amount of bioactive APO2L/TRAIL associated with exosomes in SF from RA patients. This study was undertaken to evaluate the effectiveness of bioactive APO2L/TRAIL conjugated with artificial lipid vesicles resembling natural exosomes as a treatment in a rabbit model of antigen-induced arthritis (AIA).Methods. We used a novel Ni 2؉-(N-5-amino-1-carboxypentyl)-iminodiacetic acid)-containing liposomal system. APO2L/TRAIL bound to liposomes was intraarticularly injected into the knees of animals with AIA. One week after treatment, rabbits were killed, and arthritic synovial tissue was analyzed.Results. Tethering APO2L/TRAIL to the liposome membrane increased its bioactivity and resulted in more effective treatment of AIA compared with soluble, unconjugated APO2L/TRAIL, with substantially reduced synovial hyperplasia and inflammation in rabbit knee joints. The results of biophysical studies suggested that the increased bioactivity of APO2L/TRAIL associated with liposomes was due to the increase in the local concentration of the recombinant protein, augmenting its receptor crosslinking potential, and not to conformational changes in the protein. In spite of this increase in bioactivity, the treatment lacked systemic toxicity and was not hepatotoxic.Conclusion. Our findings indicate that binding APO2L/TRAIL to the liposome membrane increases its bioactivity and results in effective treatment of AIA.

Therapeutic biomaterials based on extracellular vesicles: classification of bio‐engineering and mimetic preparation routes

“…Successful conjugation of peptides/proteins with liposomes can be checked using conventional molecular biology techniques such as dot-blot [15], SDS-PAGE [102] or even flow cytometry [102]. A preliminary purification step is required in order to remove unconjugated biomolecules.…”

“…A preliminary purification step is required in order to remove unconjugated biomolecules. For this purpose, the authors have used classical separation methods, such as ultracentrifugation [102] or gel filtration [15,103] (SEC) with high exclusion limit resins (Sepharose CL-2B, 4B mainly). Dialysis, however, is not used due to the high molecular weight of biomolecules selected for mimicking exosomes.…”

“…Martínez-Lostao et al [102] had a formulation that included lipids and stoichiometry inspired in natural exosomes. The introduction of only 5% (w/w) of an iminodiacetic acid derivative or DOGS-NTA allowed the binding of APO2L/TRAIL-His 10 to liposomes in a single step.…”

Rheumatoid Arthritis: Laboratory Models of the Disease

“…Выбор данной модели обусловлен ее сходством с человеческим РА (фибротическое разрастание синовия с формированием панну-са). Информация, почерпнутая из подобных ис-следований, может пролить свет на патогенез как раннего, так и «установившегося» артрита [33]. У кроликов, в отличие от мелких живот-ных, возможно качественное иммуногистохими-ческие исследование суставного хряща.…”

The Application of Membrane Vesicles for Cancer Therapy

“…GPA 33*, CEACAM5*, EFNB1* annexins, ARFs*, Rabs*, ADAM10*, CD44*, NG2 ephrin-B1, MIF*, bcatenin, junction plakoglobin galectin-4, RACK1*, and tetraspanin-8, FASL* AND TRAIL* FASLG* TNFSF10* (Andreola et al, 2002;Abusamra et al, 2005b;Huber et al, 2005;Taylor & Gercel-Taylor, 2005;Choi et al, 2007;Simpson et al, 2009) (Andre et al, 2002b;Mears et al, 2004;Abusamra et al, 2005b) Mesothelioma PLVAP* Ovarian ERBB2* (Andre et al, 2002b;Andreola et al, 2002;Koga et al, 2005) Prostate FASLG* TNFSF10* (Huber et al, 2005;Kim et al, 2005) Squamous cell cancer FASLG TNFSF10 (Taylor et al, 2006;Martinez-Lostao et al, 2010) Recent studies have shown that TEX provide a protective role to the cancer cells, which can be manifested in different ways. Survivin, a member of the inhibitor of apoptosis (IAP) protein family, was found to be released from tumor cells via exosomes (Khan et al, 2011).…”