Isopentenyl Pyrophosphate (IPP)

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Short Introduction of IPP

IPP is an intermediate in the classical β-Hydroxy β-methylglutaryl-CoA (HMG-CoA) reductase pathway (commonly called the mevalonate pathway) and in the non-mevalonate (methylerythritol 4-phosphate) MEP pathway of isoprenoid precursor biosynthesis. IPP toxicity presents a challenge in engineered microbial systems since its formation is unavoidable in terpene biosynthesis.

Isoprenol pathway reactions and associated heterologous genes. Fig.1 Isoprenol pathway reactions and associated heterologous genes. (Kevin, 2018)

IPP and γδ T Cell

γδ T cell receptor (TCR)-positive T cells, which control the innate immune system, display anti-tumor immunity as well as other non-immune-mediated anti-cancer effects. Nitrogen-containing bisphosphonate (N-BP) treatment expands γδ T cells ex vivo and these expanded cells can kill tumor cells in a major histocompatibility complex (MHC)-unrestricted manner. N-BP inhibits farnesyl pyrophosphate synthase in the mevalonate pathway, resulting in the accumulation of IPP, which is a stimulatory antigen for γδ T cells.

  • IPP Increased the Speed of γδ T Cell Migration

By counting the number of γδ T cells attracted across the transwell membrane to the lower chamber containing IPP, IPP induced γδ T cell migration in a dose-dependent manner can be seen.

The movement of γδ T cell migration and tracks traced by IPP and the solvent control were recorded to estimate the chemotactic indices of γδ T cells. When the solvent control was injected, γδ T cells moved at random. The γδ T cells moved toward the microinjector when IPP was introduced like the treatment of RPMI8226-CM supernatant, and the numbers of cells whose electric cell-substrate impedance sensing (ECIs) were increased by the injection of IPP. When IPP was injected, the γδ T cells moved further toward the injector than the cells treated with solvent control. These findings indicate that IPP significantly enhances the chemotactic migration of γδ T cells.

Quantitative analysis of γδ T cell migration induced by RPMI8226-conditioned medium (CM). Fig.2 Quantitative analysis of γδ T cell migration induced by RPMI8226-conditioned medium (CM). (Eishi, 2015)

  • Low Doses of Ethanol Enhances the Proliferation and Expansion of IPP Stimulated γδ T Cells

Carboxyfluorescein succinimidyl ester (CFSE)-stained synovial fluid mononuclear cells (SFMCs) from patients with different arthritic diagnoses were stimulated in vitro with IPP and IPP in combination with ethanol. The proportion of viable CFSE low proliferating γδ T cells was measured by flow cytometry. IPP, 0.25 mM, in combination with 0.015% ethanol gave a significantly higher fractional expansion, compared with IPP alone after 7 days of SFMC culture.

Expansion and proliferation of γδ T cells stimulated with IPP and ethanol for 7 days. Fig.3 Expansion and proliferation of γδ T cells stimulated with IPP and ethanol for 7 days. (Laurent, 2014)

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  1. Kevin, W.; et al. Integrated analysis of isopentenyl pyrophosphate (IPP) toxicity in isoprenoid-producing Escherichia coli. Metabolic Engineering. 2018, 47.
  2. Eishi, Ashihara.; et al. Isopentenyl pyrophosphate secreted from Zoledronate-stimulated myeloma cells, activates the chemotaxis of γδ T cells. Biochemical and Biophysical Research Communications. 2015, 463 (4).
  3. Laurent, Agneta.; et al. Synergistic effects of ethanol and isopentenyl pyrophosphate on expansion of γδ T cells in synovial fluid from patients with arthritis. PloS one. 2014, 9 (8).
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