Manufacturing and Preclinical Toxicity of GLP Grade Gene Deleted Attenuated Leishmania Donovani Parasite Vaccine

Overview

Journal Sci Rep

Specialty Science

Date 2024 Jun 25

PMID 38918456

Authors

Kumar Avishek

Mirza A Beg

Kavita Vats

Avinash Kumar Singh

Ranadhir Dey

Kamaleshwar P Singh

Rajesh Kumar Singh

Sreenivas Gannavaram

V Ramesh

Mohmad Sadik A Mulla

Upendra Bhatnagar

Sanjay Singh

Hira L Nakhasi

Poonam Salotra

Angamuthu Selvapandiyan

Affiliations

Soon will be listed here.

Abstract

Centrin1 gene deleted Leishmania donovani parasite (LdCen1) was developed and extensively tested experimentally as an intracellular stage-specific attenuated and immunoprotective live parasite vaccine candidate ex vivo using human PBMCs and in vivo in animals. Here we report manufacturing and pre-clinical evaluation of current Good-Laboratory Practice (cGLP) grade LdCen1 parasites, as a prerequisite before proceeding with clinical trials. We screened three batches of LdCen1 parasites manufactured in bioreactors under cGLP conditions, for their consistency in genetic stability, attenuation, and safety. One such batch was preclinically tested using human PBMCs and animals (hamsters and dogs) for its safety and protective immunogenicity. The immunogenicity of the CGLP grade LdCen1 parasites was similar to one grown under laboratory conditions. The cGLP grade LdCen1 parasites were found to be safe and non-toxic in hamsters and dogs even at 3 times the anticipated vaccine dose. When PBMCs from healed visceral leishmaniasis (VL) cases were infected with cGLP LdCen1, there was a significant increase in the stimulation of cytokines that contribute to protective responses against VL. This effect, measured by multiplex ELISA, was greater than that observed in PBMCs from healthy individuals. These results suggest that cGLP grade LdCen1 manufactured under cGMP complaint conditions can be suitable for future clinical trials.

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