Skip to main content

 

 

Phone: (719) 264-6250
email: info@hemogenix.com

 


Immunotoxicity 

 

Immunotoxicity testing summary performed at HemoGenix

 

For all immunotoxicity testing services please contact HemoGenix® at info@hemogenix.com, or call (719) 264-6250. Our immunotoxicity services are also available though Science Exchange.

 

The lymphopoietic and hematopoietic systems are joined at a very primitive stage of blood stem cell development. One of the cells that is common to both lineages is the SC-HPP or Stem Cell - High Proliferative Potential. This cells is quiescent, but can be affected by small molecules, which take effect once the SC-HPP is called upon to proliferate. Also affected would be the lymphopoietic progenitor cells, namely the P-Tcell and P-Bcell. 

 

Toxicity to these very primitive and early cells can be predicted using the HALO®-Tox HT and/or HALO® Real Time Platforms, but may also include colony-forming cell (CFC) assays.

  

In vitro immunotoxicity testing proper, usually begins once the T- and B-cells have been determined and demonstrate the functional characteristics of immune cells. This stage of immunotoxicity testing includes a number of functional tests that include, but not limited to, the following:

  • Lymphocyte proliferation/cytotoxicity assays 
  • Mixed lymphocyte culture/reaction (MLC/MLR)
  • Regulatory T-cell (Tregs) response
  • Cytotoxic T-cell (CTL) response
  • Antibody-dependent cell-mediated cytotoxicity (ADCC)
  • Antibody-drug congugates (ADC)
  • Immunophenotyping
  • Macrophage (M1/M2) response
  • Phagocytosis, oxidative burst (neutrophils), and migration
  • NK activity 
  • Dendritic cell maturation and co-stimulation
  • Cytokine/chemokine production and release
  • Cell injury and cell death
  • Extracellular markers of activation

 

The assays above can be performed using:

  • Peripheral blood mononuclear cells (PBMC)
  • Bone marrow (BM)
  • Purified lymphocyte cell populations

 

Species include:

  • Human
  • Primate
  • Dog
  • Rat
  • Mouse
  • Not all of these species may be available for particular studies

  • ImmunoGlo™-Tox HT is a lytic, high throughput, ATP bioluminescence platform used for screening compounds. It uses the same standardized readout as HALO®-Tox HT, but different lymphocyte culture conditions.
  • ImmunoGlo™ Real Time is a non-lytic, bioluminescence real time readout to determine the onset of immunotoxicity in a dose-dependent manner and can be simultaeously multiplexed with flow cytometric phenotyic analysis to determine which cell populations are affected by the insult.
  • ImmunoGlo™ MLC is a 1- or 2-way mixed lymphocyte reaction (MLR) or culture (MLC) in which the stimulator cells can be treated with mitomycin-C (as opposed to radiation) to inhibit their proliferation in a 1-way MLC.
  • ImmunoGlo™ TCP are assays in which co-stimulators, such as CD3 and CD28, are used to stimulate, detect and predict immunotoxicity.


Immunotoxicity Testing and Mechanism of Action (MOA) Panel 

Assay Name Assay Type Pathway Readout
ImmunoGlo™-Tox HT Standardized iATP

T- and B-Cell Proliferation

(Tier 2)

Bioluminescence
ImmunoGlo™ TCP Standardized iATP

T-cell proliferation with

co-stimulators

(Tier 2)

Bioluminescence
ImmunoGlo™-Tox Real Time (RT) Reduction potential

T- and B-Cell Proliferation

(Tier 2)

Bioluminescence
ImmunoGlo™-MLC Standardized iATP

T- and B-Cell Proliferation

(Tier 2)

Bioluminescence
Cell cycle DNA marker Proliferation (Tier 2) Fluorescence
FlowDiff™ Immune membrane expression markers (e.g. CD3, 4, 8, 19, 73) Characterization / Differentiation (Tier 2) Fluorescence
LIVEGlo™ iATP Metabolic viability / Mitochondrial Function (Tier 1 & 2) Bioluminescence
Dye Exclusion Viability 7-Aminoactinomycin D (7-AAD) / Propidium Iodide (PI) Membrane integrity (Tier 1 & 2) Fluorescence
Mitochondrial ToxGlo™* ATP Mitochondrial dysfunction (Tier 2) Bioluminescence / Fluorescence
Glutathione Assay Glutathione Oxidative stress (Tier 2) Luminescence
OxyFLOW™ 8-Oxoguanine addudcts Oxidative DNA damage (Tier 2) Fluorescence
CaspaseGlo™* Apoptosis Caspases (Tier 2) Luminescence
Annexin-V / PI Apoptosis / Necrosis Phosphatidylcholine (Tier2) Fluorescence
GFkine™  Growth factor, cytokine production / release Multiple (Tier 2) Multiple

* Promega Corporation assays 

 

For more MOA studies, please navigate to the MOA page.