Flow cytometry analysis with a new FITC-conjugated monoclonal antibody-3E12 for HLA-B*57:01 rapid screening in prevention of abacavir hypersensitivity in HIV-1-infected patients.
BACKGROUND
Rapid screening for the detection of HLA-B*57:01 in the prevention of abacavir hypersensitivity in HIV-1-infected patients is a hallmark for clinical services.


OBJECTIVE
The aim of this work was to analyze the utility of flow cytometry with a new FITC-conjugated B-17 monoclonal antibody (mAb3E12) for HLA-B*57:01 screening in a Spanish cohort of 577 HIV-1+ individuals.


METHODS
Cryopreserved peripheral blood mononuclear cell samples from HIV-1+ individuals were analyzed by flow cytometry with the mAb 3E12 that recognizes both HLA-B*57 and HLA-B*58 alleles (members of the group specificity, HLA-B17). Patients' DNA samples had been previously typed for HLA-B*57:01 with PCR-SSO or PCR-SSP and additional DNA sequencing (EPI Study). The results obtained by flow cytometry were compared with the results obtained by the DNA-PCR techniques.


RESULTS
By flow cytometry, 46 samples (7.97%) were positive for HLA-B17, 530 (91.86%) were negative, and 1 (0.17%) was undetermined. All samples found negative by flow cytometry were negative for HLA-B*57:01 by DNA-PCR. Of the HLA-B17 positive samples, 31 (67.4%) were positive for HLA-B*57:01, 2 (3.25%) were positive for HLA-B*57:03, 11 (26.1%) were positive for HLA-B*58, and 2 (3.25%) were negative for both HLA-B*57 and HLA-B*58 antigens. The undetermined sample was negative for HLA-B*57 and HLA-B*58 alleles by DNA-PCR.


CONCLUSIONS
This study shows that flow cytometry with mAb3E12 is a highly sensitive method (no false negatives) to implement prior to DNA-PCR analysis for rapid screening of HLA-B*57:01. Additional confirmation by molecular HLA typing method would be required in less than 10% of the cohort of HIV-1-infected individuals.
Servicio de Inmunología Clínica, Hospital General Universitario Gregorio Marañón, Departamento de Microbiología I (Inmunología), Universidad Complutense de Madrid, Madrid, Spain. crodriguezs.hgugm@salud.madrid.org
C
C
Rodríguez-Sáinz
L
L
Valor
D C
DC
Hernández
J
J
Gil
J
J
Carbone
M
M
Pascual-Bernaldez
F
F
Rodríguez-Alcántara
I
I
Martínez
J L
JL
Vicario
S
S
Mallal
E
E
Fernández-Cruz
D000163:Acquired Immunodeficiency Syndrome / Q000188:drug therapy*; D019380:Anti-HIV Agents / Q000009:adverse effects*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015224:Dideoxynucleosides / Q000009:adverse effects*; D004342:Drug Hypersensitivity / Q000517:prevention & control*; D005189:False Positive Reactions; D005434:Flow Cytometry / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate; D015497:HIV-1; D015235:HLA-B Antigens / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D013997:Time Factors; D000099227:Abacavir
D019380:Anti-HIV Agents; D000911:Antibodies, Monoclonal; D015224:Dideoxynucleosides; D016650:Fluorescein-5-isothiocyanate; D015235:HLA-B Antigens; D000099227:Abacavir; C103707:HLA-B57 antigen