Enumeration of functionally active anti-Aspergillus T-cells in human peripheral blood.
Invasive aspergillosis remains a life-threatening complication in patients undergoing allogeneic stem cell transplantation (SCT). Since CD4(+) T-cells provide a critical secondary defense against Aspergillus spp., the quantification of "functional" anti-Aspergillus T-cells might be important in the clinical care of allogeneic transplant patients. We present a rapid, simple and reproducible method to enumerate functionally active, cytokine-producing anti-Aspergillus T-cells in peripheral blood by means of flow cytometry, by which these cells were also phenotypically characterized as memory CD4(+) T-cells. When using 100,000 PBMCs and requiring a minimum of 50 events, at least one anti-Aspergillus T-cell among 1000 CD4(+) T-cells can be detected. Compared to healthy individuals, the number of anti-Aspergillus T-cells in patients up to one year after SCT was significantly lower. The presented method might help to define hematopoietic transplant recipients who will benefit from adoptive transfer of anti-Aspergillus T cells.
Beck
Olaf
O
Pediatric Hematology and Oncology, Children's Hospital III, Johann Wolfgang Goethe University, Frankfurt, Germany.
Koehl
Ulrike
U
Tramsen
Lars
L
Mousset
Sabine
S
Latgé
Jean Paul
JP
Müller
Katrin
K
Schwabe
Dirk
D
Bader
Peter
P
Klingebiel
Thomas
T
Lehrnbecher
Thomas
T
D000293:Adolescent; D000328:Adult; D000946:Antigens, Fungal; D001228:Aspergillosis / Q000209:etiology / Q000276:immunology* / Q000382:microbiology; D001232:Aspergillus fumigatus / Q000276:immunology*; D018791:CD4 Lymphocyte Count; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000382:microbiology; D016022:Case-Control Studies; D002478:Cells, Cultured; D002648:Child; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects*; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping / Q000379:methods*; D008297:Male; D010641:Phenotype; D015203:Reproducibility of Results; D013997:Time Factors
D000946:Antigens, Fungal; D016207:Cytokines