EPSTEIN-BARR VİRÜSÜNE KARŞI UMUT VADEDEN LİPOZOMAL AŞI ADAYI: İN VİTRO DEĞERLENDİRME

Öz

Amaç: Epstein-Barr  virüsü (EBV), enfeksiyöz  mononükleoz,  çeşitli  maligniteler  ve  otoimmün hastalıklarla ilişkili yüksek prevalansa sahip onkojenik bir herpesvirüstür. Küresel hastalık yüküne rağmen henüz lisanslı bir aşı bulunmamaktadır. Bu çalışma, tam uzunlukta gp350 antijenini MPLA adjuvanı ile enkapsüle eden PEGile lipozomal formülasyonların geliştirilmesini ve lipid bileşiminin EBV alt birim aşılarının fizikokimyasal özellikleri, biyouyumluluğu, stabilitesi ve immünomodülatör potansiyeli üzerindeki etkisini değerlendirmeyi amaçlamıştır.

Gereç ve Yöntem: Fizikokimyasal özellikleri karşılaştırmak amacıyla DOPC ve DOPE esaslı çift tabakalar kullanılarak PEGile lipozomlar hazırlanmıştır. Partikül boyutu, PDI ve zeta potansiyeli DLS yöntemiyle analiz edilmiştir. 4°C’de 30 gün boyunca stabilite ve 37°C’de antijen salım kinetiği değerlendirilmiştir. Sitotoksisite (MTT, LDH testleri), oksidatif ve genotoksik stres (ROS, comet testleri) ile THP-1 kaynaklı makrofajlarda sitokin yanıtları (TNF-α, IL-6, IL-10) incelenerek güvenlik ve immün uyarıcı potansiyel belirlenmiştir.

Sonuç ve Tartışma: Her iki formülasyon da 200 nm’den küçük, yüksek enkapsülasyon verimliliğine (>%90) sahip ve en az 30 gün boyunca stabil özellik gösteren nanopartiküller üretmiştir. DOPC esaslı lipozomlar kontrollü ve sürdürülebilir antijen salımı (24 saatte %25) sağlarken, DOPE esaslı lipozomlar hızlı salım profili (6 saat içinde %60) göstermiştir. Sitotoksisite, ROS ve comet analizleri DOPC bazlı taşıyıcının daha yüksek biyouyumluluğa sahip olduğunu ortaya koymuştur. İmmünolojik analizler, DOPC formülasyonunun güçlü TNF-α ve IL-6 ekspresyonu, orta düzeyde IL-10 salımı ve Th1 ağırlıklı bir yanıt ile dengeli bir sitokin profili indüklediğini; buna karşın DOPE esaslı formülasyonun IL-6 baskın bir yanıt oluşturduğunu göstermiştir. Bu bulgular, DOPC bazlı PEGile lipozomları gp350 antijeni için güvenli, stabil ve immünostimülatör bir taşıyıcı platform olarak tanımlamakta ve gelecek nesil EBV aşıları için umut verici bir aday olduklarını vurgulamaktadır.

Anahtar Kelimeler

• Alt birim aşısı
• Epstein–Barr virüsü
• gp350 antijeni
• lipozom
• MPLA

A PROMISING LIPOSOMAL VACCINE CANDIDATE AGAINST EPSTEIN-BARR VIRUS: AN IN VITRO EVALUATION

Öz

Objective: In this study, we developed PEGylated-liposomes encapsulating full-length gp350 with MPLA adjuvant, systematically modulating DOPC- versus DOPE-based bilayers to reveal how lipid composition influences physicochemical properties, cytotoxicity, ROS generation, genotoxicity, stability, and cytokine induction, establishing a novel nano-encapsulation platform that informs the rational design of next-generation EBV subunit vaccines capable of eliciting coordinated innate and early adaptive immune responses.

Material and Method: PEGylated-liposomes were prepared using DOPC- and DOPE-based bilayers to compare their physicochemical characteristics. Particle size, PDI, and zeta potential were analyzed by DLS. Encapsulation efficiency, stability at 4°C for 30 days, and antigen release kinetics at 37°C were determined. Cytotoxicity (MTT, LDH assays), oxidative and genotoxic stress (ROS, comet assays), and cytokine responses (TNF-α, IL-6, IL-10) in THP-1–derived macrophages were evaluated to assess safety and immunostimulatory capacity.

Result and Discussion: Both formulations produced nanoparticles smaller than 200 nm with high encapsulation efficiency (>90%) and maintained stability for at least 30 days. DOPC-based liposomes exhibited a sustained antigen release (25% at 24 h), whereas DOPE-based liposomes showed a rapid burst release (60% within 6 h). Cytotoxicity, ROS, and comet assays indicated superior biocompatibility of the DOPC-based formulation. Immunological analysis revealed that the DOPC formulation induced a balanced cytokine profile with robust TNF-α and IL-6 expression, moderate IL-10 secretion, and a Th1-skewed response, while the DOPE-based formulation displayed an IL-6–dominant pattern. Collectively, these results identify DOPC-based PEGylated-liposomes as a safe, stable, and immunostimulatory platform for gp350 delivery, highlighting their promise as a next-generation EBV vaccine candidate.

Anahtar Kelimeler

• Epstein–Barr virus
• gp350 antigen
• liposome
• MPLA
• subunit vaccine

Kaynakça

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