Table_1_Improvement of cell suspension cultures of transformed and untransformed Carica papaya cell lines, towards the development of an antiparasitic product against the gastrointestinal nematode Haemonchus contortus.pdf

Parasitic diseases have a major impact on human and animal health worldwide. Despite the availability of effective anti-parasitic drugs, their excessive and uncontrolled use has promoted the emergence of drug resistance, severely affecting ecosystems and human health. Thus, developing environmentally friendly antiparasitic treatments is urgently needed. Carica papaya has shown promising effects against infectious diseases. C. papaya embryogenic calluses were genetically modified by our research team to insert immunogenic peptides with the goal of developing an oral anti-cysticercosis vaccine. Among these callus cell lines, one labeled as CF-23, which expresses the KETc7 immunogenic peptide, induced the highest protection levels against experimental cysticercosis. In the process of designing a natural antiparasitic product based on C. papaya that simultaneously induced immunity against cysticercosis, both transformed (SF-23) and untransformed (SF-WT) suspension cultures were produced and optimized. Our results showed a better duplication time (td) for SF-23 (6.9 days) than SF-WT (13.02 days); thus, the SF-23 line was selected for scale-up in a 2-L airlift bioreactor, reaching a td of 4.4 days. This is the first time that a transgenic line of C. papaya has been grown in an airlift bioreactor, highlighting its potential for scale-up cultivation in this type of reactor. Considering the previously reported nematocidal activity of C. papaya tissues, their activity against the nematode Haemonchus contortus of aqueous extracts of SF-WT and SF-23 was explored in this study, with promising results. The information herein reported will allow us to continue the cultivation of the transgenic cell suspension line of C. papaya under reproducible conditions, to develop a new anti-parasitic product.

全球范围内，寄生虫疾病对人类和动物健康产生了重大影响。尽管有效的抗寄生虫药物已经可用，但它们过度且无控制的滥用催生了药物耐药性的产生，严重影响了生态系统和人类健康。因此，迫切需要开发环保型抗寄生虫治疗方法。木瓜（Carica papaya）在对抗传染病方面展现出良好的效果。本研究团队通过基因改造技术，将免疫原性肽插入木瓜胚胎愈伤组织中，旨在开发一种口服囊虫病疫苗。在这些愈伤细胞系中，标记为CF-23的细胞系表达KETc7免疫原性肽，诱导了对实验性囊虫病最高的保护水平。在设计基于木瓜的自然抗寄生虫产品的同时，引发了针对囊虫病的免疫反应，我们生产并优化了转化（SF-23）和非转化（SF-WT）的悬浮培养。我们的结果显示，SF-23的复制时间（td）比SF-WT短（分别为6.9天和13.02天）；因此，SF-23系被选用于2升气升式生物反应器中的扩大培养，其td降至4.4天。这是首次在气升式生物反应器中种植木瓜转基因系，突显了其在该类型反应器中扩大培养的潜力。鉴于先前报道的木瓜组织具有杀线虫活性，本研究还探讨了SF-WT和SF-23的水提取物对线虫Haemonchus contortus的活性，结果同样令人鼓舞。本报告中所述信息将使我们能够在可复制的条件下继续培养木瓜转基因细胞悬浮系，以开发一种新型抗寄生虫产品。