Skip to main content

Assessment of monocytic HLA-DR expression in ICU patients: analytical issues for multicentric flow cytometry studies

We read with interest the recent report by Gogos and colleagues [1]. While the rationale for their study is excellent, we would like to comment on technical issues that may have influenced the results.

As stated, a time limit of 8 hours between sample drawing and staining at a central laboratory was specified [1]. Unfortunately, information regarding transport conditions is missing (average time, temperature). This seems important since monocytic HLA-DR expression (mHLA-DR) increases artificially over time [2, 3]. Consequently, recommendations suggest that sample staining for mHLA-DR should occur within 4 hours [2, 3]. Although the authors aimed to address the effect of transportation, they inappropriately used samples presenting with already near-maximal mHLA-DR values (> 90%) before storage. We therefore assume that mHLA-DR results may be falsely elevated due to prolonged transportation times. Furthermore, mHLA-DR modulation during sepsis takes days and consecutive measurements are required [4]. Assessment of one early sample (within the first 24 hours) is probably inappropriate to investigate the impact of infection on mHLA-DR. Similarly, apoptosis staining should not be performed after 8 hours and experts' recommendations highlight the need for dedicated protocols on fresh cells [5].

We are convinced that successful future trials in sepsis will rely on our capacity to accurately assess immune responses. In that sense, flow cytometry multicentric clinical studies are essential. Such trials should be performed in standardized environments in accordance with specific (pre)analytical requirements. Otherwise, results might be misinterpreted and may impede promising new avenues in future care of septic patients.

Abbreviations

ICU:

intensive care unit

mHLA-DR:

monocytic human leukocyte antigen DR-1.

References

  1. 1.

    Gogos C, Kotsaki A, Pelekanou A, Giannikopoulos G, Vaki I, Maravitsa P, Adamis S, Alexiou Z, Andrianopoulos G, Antonopoulou A, Athanassia S, Baziaka F, Charalambous A, Christodoulou S, Dimopoulou I, Floros I, Giannitsioti E, Gkanas P, Ioakeimidou A, Kanellakopoulou K, Karabela N, Karagianni V, Katsarolis I, Kontopithari G, Kopterides P, Koutelidakis I, Koutoukas P, Kranidioti H, Lignos M, Louis K, et al.: Early alterations of the innate and adaptive immune statuses in sepsis according to the type of underlying infection. Crit Care 2010, 14: R96. 10.1186/cc9031

    PubMed Central  Article  PubMed  Google Scholar 

  2. 2.

    Monneret G, Elmenkouri N, Bohe J, Debard AL, Gutowski MC, Bienvenu J, Lepape A: Analytical requirements for measuring monocytic human lymphocyte antigen DR by flow cytometry: application to the monitoring of patients with septic shock. Clin Chem 2002, 48: 1589-1592.

    CAS  PubMed  Google Scholar 

  3. 3.

    Döcke WD, Höflich C, Davis KA, Röttgers K, Meisel C, Kiefer P, Weber SU, Hedwig-Geissing M, Kreuzfelder E, Tschentscher P, Nebe T, Engel A, Monneret G, Spittler A, Schmolke K, Reinke P, Volk HD, Kunz D: Monitoring temporary immunodepression by flow cytometric measurement of monocytic HLA-DR expression: a multicenter standardized study. Clin Chem 2005, 51: 2341-2347. 10.1373/clinchem.2005.052639

    Article  PubMed  Google Scholar 

  4. 4.

    Monneret G, Lepape A, Voirin N, Bohe J, Venet F, Debard AL, Thizy H, Bienvenu J, Gueyffier F, Vanhems P: Persisting low monocyte human leukocyte antigen-DR expression predicts mortality in septic shock. Intensive Care Med 2006, 32: 1175-1183. 10.1007/s00134-006-0204-8

    Article  PubMed  Google Scholar 

  5. 5.

    Ayala A, Perl M, Venet F, Lomas-Neira J, Swan R, Chung CS: Apoptosis in sepsis: mechanisms, clinical impact and potential therapeutic targets. Curr Pharm Des 2008, 14: 1853-1859. 10.2174/138161208784980617

    CAS  Article  PubMed  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Guillaume Monneret.

Additional information

Competing interests

The authors declare that they have no competing interests.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Monneret, G., Venet, F., Meisel, C. et al. Assessment of monocytic HLA-DR expression in ICU patients: analytical issues for multicentric flow cytometry studies. Crit Care 14, 432 (2010). https://doi.org/10.1186/cc9184

Download citation

Keywords

  • Septic Patient
  • Standardize Environment
  • Analytical Requirement
  • Future Care
  • Consecutive Measurement