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TruCulture

Flu Antigen Recall Responses using TruCulture® with OptiMAP and SIMOA® Analysis

Shen-An Hwang, Dominic Eisinger, and Samuel LaBrie

ABSTRACT

TruCulture® is a whole blood collection and culture system that is designed to be used at clinical sites to investigate human immune responses under specific stimulation conditions. OptiMAP is a Multi-Analyte Profile (MAP) bead based immunoassay panel that measures 13 analytes that span the major immune responses (TH1, TH2, TH17, neutrophil, and monocyte/macrophage activation). The recall response of flu hemagglutinin (HA) antigens was examined in healthy adults vaccinated with the 2018-2019 annual flu vaccine. Whole blood was drawn into TruCulture tubes: null (media only), staphylococcal enterotoxin B (SEB), and null tubes spiked with 1.25mg each of 4 different recombinant HA proteins. All samples were incubated for 48 hours at 37oC. Supernatant were collected and analyzed using the OptiMAP panel. No inflammatory cytokines were detected in the null tubes from either the baseline or 2 weeks post-vaccination samples. There were no differences in cytokine production in the SEB stimulated samples between the baseline and 2 weeks post-flu vaccination time lines. For flu HA stimulated samples, there was a significant increase in production of GM-CSF, IFN-γ, IL-1β, IL-6, IL-12p70, IL-23, and TNF-α at 2 weeks post-flu vaccination compared to samples collected at baseline. Additionally, flu HA stimulated samples post vaccination also demonstrated increased production of type I inteferons, as analyzed by SIMOA. These data demonstrate that TruCulture in conjunction with OptiMAP and SIMOA are useful tools for investigating antigen specific responses.

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How TruCulture Works

TruCulture tubes are pre-loaded with cell culture media and immune stimulant(s) or drug candidates. Blood is drawn directly into the TruCulture tube and incubated in a dry heat block. Supernatants are collected by simply inserting a valve separator to separate cells from the culture supernatant.

TruCulture Procedure, Collect

01. COLLECT

Draw 1 mL of blood directly into the TruCulture Tube and break off the plunger.

TruCulture Procedure, MIX

02. MIX

Gently invert tube to mix 3 to5 times

TruCulture Procedure, Incubate

02. INCUBATE

Place tube in 37ºC heat block for up to 24 or 48 hours.

TruCulture Procedure, separate

02. SEPARATE

Manually insert valve to separate supernatant from the cells. Collect supernatant and cell layer for downstream analysis.

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Applications

TruCulture has been utilized as a whole-blood stimulation system by researchers and drug developers in several fields to reliably measure immune response for the following applications.

  • Pharmacodynamics (including dose response)
  • Functional immune cell analysis
  • Disease characterization
  • Patient stratification
  • Genotype – to – Phenotype association studies
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PBMCs vs. TruCulture

Traditional pharmacodynamic whole blood experiments are generally short in duration (2-6 hrs) as a consequence of poor culture conditions leading to premature termination of the normal physiological immune response. A longer more robust response usually provides higher sensitivity and greater relevance than short incubation times that may only lead to the release of stored pre-synthesized mediators.

Advantages of TruCulture

  • Integrated closed sterile instant whole blood collection and culture system.
  • Standardized to ensure consistent performance across multiple users and clinical sites.
  • Reliable, easy to use, and reproducible – eliminates the need for cell manipulation.
  • Retains all blood components, granulocytes, platelets, red blood cells, soluble factors and Fc receptor expressing cells.
  • Only inexpensive heat block needed, no lab equipment nor centrifugation steps.
  • Has been successfully deployed in hundreds of clinical drug trials.

Disadvantages of PBMCs

  • Separate blood collection and specialized cell culture procedures.
  • Extensive manipulation, processing, and often freezing/shipping prior to culturing.
  • Requires technical expertise with increased variability across users and clinical sites.
  • Requires CO2 incubator, biosafety cabinet, centrifuge, media, and cell culture plastics.
  • Culture procedures/conditions are difficult to standardize for clinical trial applications.
  • Open, less sterile, artificial system.
  • Poor reproducibility.
PBMC vs TruCulture whole blood collection and stimulation

Learn why TruCulture Whole Blood Culture System is a more relevant model of human leukocyte function than the traditional method of cultured PBMCs.

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