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Lactococcus lactis expressing sand fly PpSP15 salivary protein confers long-term protection against Leishmania major in BALB/c mice.
PLOS Neglected Tropical Diseases ( IF 3.4 ) Pub Date : 2020-01-03 , DOI: 10.1371/journal.pntd.0007939 Elaheh Davarpanah 1, 2 , Negar Seyed 2 , Fariborz Bahrami 3 , Sima Rafati 2 , Reza Safaralizadeh 1 , Tahereh Taheri 2
PLOS Neglected Tropical Diseases ( IF 3.4 ) Pub Date : 2020-01-03 , DOI: 10.1371/journal.pntd.0007939 Elaheh Davarpanah 1, 2 , Negar Seyed 2 , Fariborz Bahrami 3 , Sima Rafati 2 , Reza Safaralizadeh 1 , Tahereh Taheri 2
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Cutaneous leishmaniasisis a vector-borne disease transmitted by Leishmania infected sand flies. PpSP15 is an immunogenic salivary protein from the sand fly Phlebotomus papatasi. Immunization with PpSP15 was shown to protect against Leishmania major infection. Lactococcus lactis is a safe non-pathogenic delivery system that can be used to express antigens in situ. Here, the codon-optimized Ppsp15-egfp gene was cloned in pNZ8121 vector downstream of the PrtP signal peptide that is responsible for expression and secretion of the protein on the cell wall. Expression of PpSP15-EGFP recombinant protein was monitored by immunofluorescence, flow cytometry and Western blot. Also, expression of protein in cell wall compartment was verified using whole cell ELISA, Western blot and TEM microscopy. BALB/c mice were immunized three times with recombinant L. lactis-PpSP15-EGFPcwa, and the immune responses were followed up, at short-term (ST, 2 weeks) and long-term (LT, 6 months) periods. BALB/c mice were challenged with L. major plus P. papatasi Salivary Gland Homogenate. Evaluation of footpad thickness and parasite burden showed a delay in the development of the disease and significantly decreased parasite numbers in PpSP15 vaccinated animals as compared to control group. In addition, immunized mice showed Th1 type immune responses. Importantly, immunization with L. lactis-PpSP15-EGFPcwa stimulated the long-term memory in mice which lasted for at least 6 months.
中文翻译：
表达白蛉 PpSP15 唾液蛋白的乳酸乳球菌可为 BALB/c 小鼠提供针对重大利什曼原虫的长期保护。
皮肤利什曼病是一种由感染利什曼原虫的沙蝇传播的媒介传播疾病。 PpSP15 是来自白蛉 Phlebotomus papatasi 的免疫原性唾液蛋白。 PpSP15 免疫被证明可以预防利什曼原虫重大感染。乳酸乳球菌是一种安全的非致病性递送系统，可用于原位表达抗原。在这里，密码子优化的 Ppsp15-egfp 基因被克隆到 pNZ8121 载体中 PrtP 信号肽的下游，该信号肽负责细胞壁上蛋白质的表达和分泌。通过免疫荧光、流式细胞术和蛋白质印迹监测PpSP15-EGFP重组蛋白的表达。此外，还使用全细胞 ELISA、Western blot 和 TEM 显微镜验证了细胞壁区室中蛋白质的表达。用重组乳酸乳球菌-PpSP15-EGFPcwa对BALB/c小鼠进行3次免疫，并对短期（ST，2周）和长期（LT，6个月）的免疫反应进行随访。 BALB/c 小鼠用 L. Major 和 P. papatasi 唾液腺匀浆攻击。足垫厚度和寄生虫负荷的评估显示，与对照组相比，PpSP15 疫苗接种动物的疾病发展延迟，寄生虫数量显着减少。此外，免疫小鼠表现出Th1型免疫反应。重要的是，用乳酸乳球菌-PpSP15-EGFPcwa 进行免疫刺激了小鼠的长期记忆，持续至少 6 个月。
更新日期：2020-01-04
中文翻译：
表达白蛉 PpSP15 唾液蛋白的乳酸乳球菌可为 BALB/c 小鼠提供针对重大利什曼原虫的长期保护。
皮肤利什曼病是一种由感染利什曼原虫的沙蝇传播的媒介传播疾病。 PpSP15 是来自白蛉 Phlebotomus papatasi 的免疫原性唾液蛋白。 PpSP15 免疫被证明可以预防利什曼原虫重大感染。乳酸乳球菌是一种安全的非致病性递送系统，可用于原位表达抗原。在这里，密码子优化的 Ppsp15-egfp 基因被克隆到 pNZ8121 载体中 PrtP 信号肽的下游，该信号肽负责细胞壁上蛋白质的表达和分泌。通过免疫荧光、流式细胞术和蛋白质印迹监测PpSP15-EGFP重组蛋白的表达。此外，还使用全细胞 ELISA、Western blot 和 TEM 显微镜验证了细胞壁区室中蛋白质的表达。用重组乳酸乳球菌-PpSP15-EGFPcwa对BALB/c小鼠进行3次免疫，并对短期（ST，2周）和长期（LT，6个月）的免疫反应进行随访。 BALB/c 小鼠用 L. Major 和 P. papatasi 唾液腺匀浆攻击。足垫厚度和寄生虫负荷的评估显示，与对照组相比，PpSP15 疫苗接种动物的疾病发展延迟，寄生虫数量显着减少。此外，免疫小鼠表现出Th1型免疫反应。重要的是，用乳酸乳球菌-PpSP15-EGFPcwa 进行免疫刺激了小鼠的长期记忆，持续至少 6 个月。
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