A second generation leishmanization vaccine with a markerless attenuated Leishmania major strain using CRISPR gene editing

• Published in: Nature communications Vol. 11; no. 1; pp. 3461 - 14
• Main Authors: Zhang, Wen-Wei, Karmakar, Subir, Gannavaram, Sreenivas, Dey, Ranadhir, Lypaczewski, Patrick, Ismail, Nevien, Siddiqui, Abid, Simonyan, Vahan, Oliveira, Fabiano, Coutinho-Abreu, Iliano V., DeSouza-Vieira, Thiago, Meneses, Claudio, Oristian, James, Serafim, Tiago D., Musa, Abu, Nakamura, Risa, Saljoughian, Noushin, Volpedo, Greta, Satoskar, Monika, Satoskar, Sanika, Dagur, Pradeep K., McCoy, J. Philip, Kamhawi, Shaden, Valenzuela, Jesus G., Hamano, Shinjiro, Satoskar, Abhay R., Matlashewski, Greg, Nakhasi, Hira L.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 10.07.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 42/41; 45/23; 45/77; 631/1647/1511; 631/250/590/1867; 631/326/417/2546; 64/60; 692/308/575; 96/31; Antibiotic resistance; Antibiotics; Clinical trials; CRISPR; Disease transmission; Drug resistance; Genetic engineering; Genetic modification; Genome editing; Genomes; Humanities and Social Sciences; Immune response; Immune system; Immunization; Leishmania; Leishmania major; Leishmaniasis; Lesions; Markers; multidisciplinary; Mutation; Parasites; Protozoa; Science; Science (multidisciplinary); Skin diseases; Skin lesions; Tropical diseases; Vaccination; Vaccines; Vector-borne diseases
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-17154-z

Leishmaniasis is a neglected tropical disease caused by Leishmania protozoa transmitted by infected sand flies. Vaccination through leishmanization with live Leishmania major has been used successfully but is no longer practiced because it resulted in occasional skin lesions. A second generation leishmanization is described here using a CRISPR genome edited L. major strain ( LmCen −/− ). Notably, LmCen −/− is a genetically engineered centrin gene knock-out mutant strain that is antibiotic resistant marker free and does not have detectable off-target mutations. Mice immunized with LmCen −/− have no visible lesions following challenge with L. major -infected sand flies, while non-immunized animals develop large and progressive lesions with a 2-log fold higher parasite burden. LmCen −/− immunization results in protection and an immune response comparable to leishmanization. LmCen −/− is safe since it is unable to cause disease in immunocompromised mice, induces robust host protection against vector sand fly challenge and because it is marker free, can be advanced to human vaccine trials. Here, the authors engineer an attenuated knock-out Leishmania ( LmCen −/− ) vaccine that is safe in immunocompromised mice and induces an immune response and protection similar to leishmanization with wild-type Leishmania . Since LmCen −/− is antibiotic resistant marker free, it is a candidate for clinical development.