Metabolic engineering of Salmonella vaccine bacteria to boost human Vγ2Vδ2 T cell immunity

Grefachew Workalemahu; Hong Wang; Kia-Joo Puan; Mohanad H Nada; Tomohisa Kuzuyama; Bradley D Jones; Chenggang Jin; Craig T Morita

doi:10.4049/jimmunol.1302746

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Metabolic engineering of Salmonella vaccine bacteria to boost human Vγ2Vδ2 T cell immunity

Journal article

Peer reviewed

Metabolic engineering of Salmonella vaccine bacteria to boost human Vγ2Vδ2 T cell immunity

Grefachew Workalemahu, Hong Wang, Kia-Joo Puan, Mohanad H Nada, Tomohisa Kuzuyama, Bradley D Jones, Chenggang Jin and Craig T Morita

The Journal of immunology (1950), Vol.193(2), pp.708-721

07/15/2014

DOI: 10.4049/jimmunol.1302746

PMCID: PMC4241231

PMID: 24943221

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Abstract

Human Vγ2Vδ2 T cells monitor isoprenoid metabolism by recognizing foreign (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), a metabolite in the 2-C-methyl-D-erythritol-4-phosphate pathway used by most eubacteria and apicomplexan parasites, and self isopentenyl pyrophosphate, a metabolite in the mevalonate pathway used by humans. Whereas microbial infections elicit prolonged expansion of memory Vγ2Vδ2 T cells, immunization with prenyl pyrophosphates or aminobisphosphonates elicit short-term Vγ2Vδ2 expansion with rapid anergy and deletion upon subsequent immunizations. We hypothesized that a live, attenuated bacterial vaccine that overproduces HMBPP would elicit long-lasting Vγ2Vδ2 T cell immunity by mimicking a natural infection. Therefore, we metabolically engineered the avirulent aroA(-) Salmonella enterica serovar Typhimurium SL7207 strain by deleting the gene for LytB (the downstream enzyme from HMBPP) and functionally complementing for this loss with genes encoding mevalonate pathway enzymes. LytB(-) Salmonella SL7207 had high HMBPP levels, infected human cells as efficiently as did the wild-type bacteria, and stimulated large ex vivo expansions of Vγ2Vδ2 T cells from human donors. Importantly, vaccination of a rhesus monkey with live lytB(-) Salmonella SL7207 stimulated a prolonged expansion of Vγ2Vδ2 T cells without significant side effects or anergy induction. These studies provide proof-of-principle that metabolic engineering can be used to derive live bacterial vaccines that boost Vγ2Vδ2 T cell immunity. Similar engineering of metabolic pathways to produce lipid Ags or B vitamin metabolite Ags could be used to derive live bacterial vaccine for other unconventional T cells that recognize nonpeptide Ags.

Immunization

Animals

Bacterial Proteins - genetics

Bacterial Proteins - immunology

Bacterial Proteins - metabolism

Cell Proliferation

Cells, Cultured

Gene Deletion

Humans

Lymphocyte Activation - immunology

Macaca mulatta - immunology

Metabolic Engineering - methods

Metabolic Networks and Pathways - genetics

Metabolic Networks and Pathways - immunology

Mevalonic Acid - metabolism

Organophosphates - metabolism

Receptors, Antigen, T-Cell, gamma-delta - immunology

Receptors, Antigen, T-Cell, gamma-delta - metabolism

Salmonella typhimurium - genetics

Salmonella typhimurium - immunology

Salmonella typhimurium - metabolism

Salmonella Vaccines - immunology

T-Lymphocytes - immunology

T-Lymphocytes - metabolism

Details

Title: Subtitle: Metabolic engineering of Salmonella vaccine bacteria to boost human Vγ2Vδ2 T cell immunity
Creators: Grefachew Workalemahu - Division of Immunology, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242; Department of Veterans Affairs, Iowa City Health Care System, Iowa City, IA 52246
Hong Wang - University of Iowa, Internal Medicine
Kia-Joo Puan - Division of Immunology, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242; Singapore Immunology Network, Agency for Science, Technology and Research, Singapore 138648
Mohanad H Nada - Division of Immunology, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242; Department of Veterans Affairs, Iowa City Health Care System, Iowa City, IA 52246; Interdisciplinary Graduate Program in Immunology, University of Iowa Carver College of Medicine, Iowa City, IA 52242
Tomohisa Kuzuyama - Biotechnology Research Center, The University of Tokyo, Tokyo 113-8657, Japan
Bradley D Jones - University of Iowa, Microbiology and Immunology
Chenggang Jin - Division of Immunology, Department of Internal Medicine, University of Iowa Carver College of Medicine, Iowa City, IA 52242; Department of Veterans Affairs, Iowa City Health Care System, Iowa City, IA 52246
Craig T Morita - University of Iowa, Internal Medicine
Resource Type: Journal article
Publication Details: The Journal of immunology (1950), Vol.193(2), pp.708-721
DOI: 10.4049/jimmunol.1302746
PMID: 24943221
PMCID: PMC4241231
NLM abbreviation: J Immunol
ISSN: 0022-1767
eISSN: 1550-6606
Grant note: I01 BX000972 / BLRD VA I01 BX000513 / BLRD VA 2P01 AI044464-2 / NIAID NIH HHS 5T32 AI007485 / NIAID NIH HHS AI057160 / NIAID NIH HHS T32 AI007485 / NIAID NIH HHS CA097274 / NCI NIH HHS P01 AI044642 / NIAID NIH HHS P30CA086862 / NCI NIH HHS P30 CA086862 / NCI NIH HHS U54 AI057160 / NIAID NIH HHS P50 CA097274 / NCI NIH HHS
Language: English
Date published: 07/15/2014
Academic Unit: Microbiology and Immunology; Immunology; Internal Medicine
Record Identifier: 9984208257202771

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