Gifre-Renom Laia, Ugarte-Berzal Estefania, Martens Erik, Boon Lise, Cano-Garrido Olivia, Martínez-Núñez Esther, Luque Teresa, Roca-Pinilla Ramon, Conchillo-Solé Òscar, Ferrer-Miralles Neus, Villaverde Antonio, Opdenakker Ghislain, Garcia-Fruitós Elena, Arís Anna
Department of Ruminant Production, Institut de Recerca i Tecnologia Agroalimentàries (IRTA), 08140 Caldes de Montbui, Spain.
Laboratory of Immunobiology, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, KU Leuven, University of Leuven, 3000 Leuven, Belgium.
Pharmaceutics. 2020 May 13;12(5):450. doi: 10.3390/pharmaceutics12050450.
Bacterial inclusion bodies (IBs) are protein-based nanoparticles of a few hundred nanometers formed during recombinant protein production processes in different bacterial hosts. IBs contain active protein in a mechanically stable nanostructured format that has been broadly characterized, showing promising potential in different fields such as tissue engineering, protein replacement therapies, cancer, and biotechnology. For immunomodulatory purposes, however, the interference of the format immunogenic properties-intrinsic to IBs-with the specific effects of the therapeutic protein is still an uncovered gap. For that, active and inactive forms of the catalytic domain of a matrix metalloproteinase-9 (MMP-9 and mutMMP-9, respectively) have been produced as IBs and compared with the soluble form for dermal inflammatory effects in knock-out mice. After protein injections in air-pouches in the mouse model, MMP-9 IBs induce local neutrophil recruitment and increase pro-inflammatory chemokine levels, lasting for at least two days, whereas the effects triggered by the soluble MMP-9 format fade out after 3 h. Interestingly, the IB intrinsic effects (mutMMP-9 IBs) do not last more than 24 h. Therefore, it may be concluded that IBs could be used for the delivery of therapeutic proteins, such as immunomodulating proteins while preserving their stability in the specific tissue and without triggering important unspecific inflammatory responses due to the protein format.
细菌包涵体（IBs）是在不同细菌宿主的重组蛋白生产过程中形成的几百纳米大小的基于蛋白质的纳米颗粒。IBs以机械稳定的纳米结构形式包含活性蛋白，这种形式已得到广泛表征，在组织工程、蛋白质替代疗法、癌症和生物技术等不同领域显示出有前景的潜力。然而，就免疫调节目的而言，IBs固有的形式免疫原性对治疗性蛋白质特定效应的干扰仍是一个未被揭示的空白。为此，基质金属蛋白酶-9（分别为MMP-9和mutMMP-9）催化结构域的活性和非活性形式已作为IBs产生，并与可溶性形式在基因敲除小鼠中进行皮肤炎症效应的比较。在小鼠模型的气袋中注射蛋白质后，MMP-9 IBs诱导局部中性粒细胞募集并增加促炎趋化因子水平，持续至少两天，而可溶性MMP-9形式引发的效应在3小时后消失。有趣的是，IBs的固有效应（mutMMP-9 IBs）持续不超过24小时。因此，可以得出结论，IBs可用于递送治疗性蛋白质，如免疫调节蛋白，同时在特定组织中保持其稳定性，且不会因蛋白质形式引发重要的非特异性炎症反应。
