False-Positive Results for Human Immunodeficiency Virus Type 1 Nucleic Acid Amplification Testing in Chimeric Antigen Receptor T Cell Therapy

Abstract

Chimeric antigen receptor (CAR) T cell immunotherapy has been a major advancement in cancer therapeutics. Reprogramming of T cells is achieved by using gammaretroviral or lentiviral vectors, which may interfere with human immunodeficiency virus type 1 (HIV-1) nucleic acid amplification testing (NAAT). Here, we describe three clinical scenarios in which CAR T cell immunotherapy interfered with HIV-1 testing, including (i) routine infectious disease screening prior to stem cell transplantation in a 16-year-old female with B cell acute lymphoblastic leukemia, post CAR T cell treatment; (ii) routine infectious disease screening prior to second CAR T cell collection in a 65-year-old male with diffuse large B cell lymphoma who failed initial CAR T cell treatment; and (iii) routine infectious risk assessment following an occupational health exposure from a 58-year-old male with multiple myeloma, who received CAR T cell treatment. In each case, patients initially tested negative by the “fourth-generation” HIV-1 screening enzyme immunoassay (targeting the p24 antigen and anti-HIV-1 antibodies), but positive by the Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test v2.0 (targeting gag and the long terminal repeat [LTR]). These samples subsequently retested negative using the Abbott m2000 RealTime HIV-1 assay, which targets the integrase gene. These results indicated that cross-reactions between lentiviral vectors and LTR genomes targeted in the HIV-1 NAAT caused the HIV-1 NAAT false-positive results.

CD19+ lineage chimerism, an early biomarker after anti-CD19 CAR-T cell therapy in patients previously receiving a hematopoietic stem cell transplantation

“…Nevertheless, our findings (patients #2 and #4) show the loss of completed chimerism observed in the CD19 + cells subset could be associated with a higher risk of relapse, even if not accompanied by a loss of CD19 + lymphocyte aplasia or a significant decrease in HIV viral load (patient #2), being the last one an indirect biomarker which could associate cross-reactions with HIV infection, leading to false-positive results ( 13 – 15 ). Although there is not enough evidence on optimal treatment in this case, our findings, supported by other publications ( 16 ), suggest that DLI may be not as effective as in other settings ( 17 ).…”

Mass Cytometry: a robust platform for the comprehensive immunomonitoring of CAR‐T‐cell therapies

“…CAR-T cells were usually assessed by flow cytometry, using the CAR Detection Reagents (Miltenyi), and by quantitative polymerase chain reaction (qPCR) using an assay commercialized to quantify HIV-1 (Generic HIV DNA Cell â test for research use, Biocentric, Bandol, France). 10,11 These two assays performed similarly (r = 0Á6782, P < 0Á0001 by Spearman correlation; n = 44; Fig 1A and Figure S1). Over a broad range of frequencies from 0Á12 to 22% CD3 + CAR-T cells, we showed that, for the detection of CAR-T cells, results obtained with our 106/116 Cd-anti-biotin CD19Fc used for mass cytometry strongly correlated with those obtained by flow cytometry (Pearson correlation; r = 0Á95, P < 0Á05; n = 72; confirmed by the validation method of Bland-Altman; Fig 1B).…”

“…Therefore, we customized the Maxpar panel by developing a cadmium ( 106/116 Cd)–anti‐biotin CD19Fc (106/116 cadmium metal isotopes, Fluidigm, Canada; pure anti‐biotin antibody and CD19Fc Detection Reagent, biotin, Milteny Biotec, Bergisch Gladbach, Germany) that identify CAR‐T cells and validated its robustness by comparison with routine assays. CAR‐T cells were usually assessed by flow cytometry, using the CAR Detection Reagents (Miltenyi), and by quantitative polymerase chain reaction (qPCR) using an assay commercialized to quantify HIV‐1 (Generic HIV DNA Cell ® test for research use, Biocentric, Bandol, France) 10,11 . These two assays performed similarly ( r = 0·6782, P < 0·0001 by Spearman correlation; n = 44; Fig 1A and Figure S1).…”

Evaluating the Dual-Target Aptima HIV-1 Quant Dx Assay: Comparison between Viral Loads Measured with pol and LTR Targets in the Same Samples

“…Recently, in HIV-1 seronegative cancer patients under chimeric antigen receptor (CAR) T-cell immunotherapy, it has been described false HIV-1 nucleic acid amplification with a double-targets assay different from Aptima ( 16 , 17 ). This was attributed to the detection of LTR elements present in lentiviral vectors used in this type of immunotherapy ( 16 , 17 ).…”