i.Mune® CD4/CD8 [RUO]


i.Mune CD4/CD8 [RUO] is a quantitative in vitro test to determine the percentages and absolute counts* of human CD4+ and CD8+ T lymphocytes in liquid venous whole blood and capillary whole blood specimens dried on filter paper (Dried Blood Spot; DBS) or Plasma Separation Cards (PSC).

(*absolute quantification in liquid blood (fresh or frozen), only)

Please note - i.Mune CD4/CD8 is for research use only. Not for diagnostic purposes.

Benefits

i.Mune CD4/CD8 [RUO] enables the epigenetic quantification of:

  • Helper T lymphocytes (CD3+CD4+)
  • Cytotoxic T lymphocytes (CD3+CD8+)

Throughput:

  • Up to 20 samples per PCR-run (96-well)

Contents

i.Mune CD4/CD8 is comprised of the three (3) separate kits:

  • i.Mune Prep (for preparation and bisulfite-conversion of DNA from fresh, frozen or dried blood samples)
  • i.Mune CD4/CD8 Amp (for PCR-amplification of bisulfite-converted DNA)
  • i.Mune CD4/CD8 Check (Positive and negative controls, standards)
  • Data Analysis Tool (MS-Excel based)

Sample Requirements


  • Capillary whole blood dried on filter paper (DBS) or Plasma Separation Cards (PSC)
    • Samples can be stored at room temperature (15°C to 30°C) or frozen (-30°C to -15°C) for up to 12 weeks
    • DBS sample stability has been demonstrated at 55°C and 65-78% humidity for up to 7 days 

OR

  • 40 µl of liquid venous whole blood collected in K2EDTA blood collection tube
    • Samples can be stored at room temperature (15°C to 30°C) for up to 24 h or frozen (-30°C to -15°C) for up to 12 weeks

Applications

Quantification of  CD4+ and CD8+ lymphocytes can be useful for*:

  • Monitoring of HIV-positive patients 1),2)
  • An increased CD4/CD8 ratio can be a sign for autoimmunity, T-cell lymphoma or multiple sclerosis 3)

*The above clinical applications have been established using technologies currently being employed in clinical laboratory routine (e.g. flow cytometry). 
Please note - i.Mune CD4/CD8 is for research use only. Not for diagnostic purposes.

Performance

Comparison i.Mune CD4/CD8 [RUO] with flow cytometry

 

247 frozen venous whole blood samples from patients with various different primary and secondary immunodeficiencies were analyzed using i.Mune CD4/CD8 and compared to flow cytometry data obtained at time of blood sampling. Correlation was determined using the Spearman correlation coefficient (Spearman r) for each assay (cells/µl and % leukocytes). 

Epigenetic quantification of CD4+ and CD8+ T-cells in Dried Blood Spots (DBS)

97 blood samples from HIV positive subjects (liquid and dried) were analyzed using flow cytometry (liquid blood) and epigenetic qPCR (liquid and dried), respectively. The data show excellent correlation between both testing methods and between liquid and dried blood 1).

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References

  1. Baron U. et al. Epigenetic immune cell counting in human blood samples for immunodiagnostics. Sci Transl Med. 2018 Aug 1; 10 (452)
  2. Ford N, et al. The evolving role of CD4 cell counts in HIV care. Curr Opin HIV AIDS. 2017 Mar;12(2):123-128
  3. https://www.gesundheit.gv.at/labor/laborwerte/immunsystem/cd43-cd83-ratio.html

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