title: A new cytokine release assay: a simple approach to monitor the immune status of HIV-infected patients.
abstract: OBJECTIVE
  To test a new assay based on an ex vivo cytokine release from whole blood for the monitoring of immune changes in human immunodeficiency virus (HIV)-infected patients.
  
  
  METHODS
  A pilot study of outpatients with HIV infection (n = 9) at a large academic hospital who were divided into three groups: HIV-infected patients on highly active antiretroviral therapy (HAART) with a CD4(+) cell count >350/μL (group I) or a CD4(+) cell count <350/μL (group II) and HIV-infected HAART-naïve subjects with a CD4(+) cell count >350/μL (group III). All groups were compared with healthy volunteers (n = 3). The ex vivo cytokine release assay was performed in a three-step process: (1) blood collection, (2) whole-blood ex vivo incubation over 48 h without or with a standard set of well-defined recall antigens as comparable to those used formerly in the skin delayed-type hypersensitivity (DTH) test, (3) cytokine determination from the assay supernatant.
  
  
  RESULTS
  Under stimulated conditions, untreated HIV-infected patients with a CD4(+) count >350/μL had similar interleukin-2 (IL-2) levels in the supernatant of the whole-blood incubation to HIV-infected patients on HAART with a low CD4(+) count. Both groups revealed lower IL-2 levels in the supernatant than HIV-infected patients on HAART and with a CD4(+) count >350/μL or healthy volunteers. The determination of interferon-γ and tumour necrosis factor-α in the supernatant showed a similar arrangement of cytokines between groups.
  
  
  CONCLUSIONS
  Our results suggest that this cytokine release assay could be a suitable tool to mirror the immunological responsiveness of patients with HIV infection in a gradual manner; further studies are required in order to assess its value in HAART monitoring.
authors.affiliation: Department of Anesthesiology, University of Munich, Marchioninistrasse 15, 81377, Munich, Germany. ines.kaufmann@med.uni-muenchen.de
authors.forename: I
authors.initials: I
authors.lastname: Kaufmann
authors.forename: R
authors.initials: R
authors.lastname: Draenert
authors.forename: M
authors.initials: M
authors.lastname: Gruber
authors.forename: M
authors.initials: M
authors.lastname: Feuerecker
authors.forename: J
authors.initials: J
authors.lastname: Roider
authors.forename: A
authors.initials: A
authors.lastname: Choukèr
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019380:Anti-HIV Agents / Q000627:therapeutic use; D023241:Antiretroviral Therapy, Highly Active / Q000379:methods; D016207:Cytokines / Q000032:analysis* / Q000378:metabolism; D016903:Drug Monitoring / Q000379:methods*; D005260:Female; D015658:HIV Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000276:immunology*; D006801:Humans; D007159:Immunologic Tests / Q000379:methods*; D008297:Male; D008875:Middle Aged
chemical_list: D019380:Anti-HIV Agents; D016207:Cytokines
references.citation: Clin Diagn Lab Immunol. 2003 Jan;10(1):133-9
references.pmid: 12522051
references.citation: Infection. 2013 Apr;41(2):545-51
references.pmid: 23264096
references.citation: Infection. 2013 Feb;41(1):41-8
references.pmid: 23292663
references.citation: Clin Exp Immunol. 2013 May;172(2):290-9
references.pmid: 23574325
references.citation: Scand J Infect Dis. 1996;28(2):125-30
references.pmid: 8792477
references.citation: J Clin Invest. 1989 Dec;84(6):1892-9
references.pmid: 2574188
references.citation: JAMA. 2012 Jul 25;308(4):387-402
references.pmid: 22820792
references.citation: Nat Rev Immunol. 2012 Feb 17;12(3):180-90
references.pmid: 22343569
references.citation: J Vis Exp. 2011 Mar 01;(49):
references.pmid: 21403634
references.citation: Nature. 2006 Jun 15;441(7095):890-3
references.pmid: 16778891
references.citation: Blood. 2013 Jan 31;121(5):801-11
references.pmid: 23233659
references.citation: Am J Respir Crit Care Med. 2012 Feb 15;185(4):427-34
references.pmid: 22161162
references.citation: Infection. 2012 Dec;40(6):627-34
references.pmid: 22791407
references.citation: Clin Infect Dis. 2009 Apr 1;48(7):954-62
references.pmid: 19245343
references.citation: Nature. 1989 Jun 1;339(6223):383-5
references.pmid: 2524668