Detection of Tumor-Specific Antigens in Philadelphia Chromosome Positive Leukemias

Denderen, J van; Hacken, P. Ten; Berendes, P; Ewijk, Willem van

doi:10.3109/10428199309047859

Cited by 8 publications

(4 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We attempted to raise antibodies against the tumorspecific fusion epitopes of the BCR-ABL proteins. Indeed, polyclonal rabbit antisera could be raised against the p190 (e1-a2), p210 (e13-a2) and against p210 (e14-a2) epitopes, [28][29][30] and even the monoclonal antibody (MoAb), ER-FP1, was made against the p190 (e1-a2) fusion epitope. 31 These antibodies worked under denaturing conditions, such as in western blotting, but were not suited for intracellular staining in microscopy or flow cytometry.…”

Section: Introductionmentioning

confidence: 99%

Flow cytometric immunobead assay for the detection of BCR–ABL fusion proteins in leukemia patients

Weerkamp

Dekking

et al. 2009

Leukemia

View full text Add to dashboard Cite

BCR-ABL fusion proteins show increased signaling through their ABL tyrosine kinase domain, which can be blocked by specific inhibitors, thereby providing effective treatment. This makes detection of BCR-ABL aberrations of utmost importance for diagnosis, classification and treatment of leukemia patients. BCR-ABL aberrations are currently detected by karyotyping, fluorescence in situ hybridization (FISH) or PCR techniques, which are time consuming and require specialized facilities. We developed a simple flow cytometric immunobead assay for detection of BCR-ABL fusion proteins in cell lysates, using a bead-bound anti-BCR catching antibody and a fluorochromeconjugated anti-ABL detection antibody. We noticed protein stability problems in lysates caused by proteases from mature myeloid cells. This problem could largely be solved by adding protease inhibitors in several steps of the immunobead assay. Testing of 145 patient samples showed fully concordant results between the BCR-ABL immunobead assay and reverse transcriptase PCR of fusion gene transcripts. Dilution experiments with BCR-ABL positive cell lines revealed sensitivities of at least 1%. We conclude that the BCR-ABL immunobead assay detects all types of BCR-ABL proteins in leukemic cells with high specificity and sensitivity. The assay does not need specialized laboratory facilities other than a flow cytometer, provides results within B4 h, and can be run in parallel to routine immunophenotyping.

show abstract

Section: Introductionmentioning

confidence: 99%

Flow cytometric immunobead assay for the detection of BCR–ABL fusion proteins in leukemia patients

Weerkamp

Dekking

et al. 2009

Leukemia

View full text Add to dashboard Cite

show abstract

“…CML cells are known to express various tumor antigens, including the bcr͞abl fusion protein, a neoantigen not shared by any normal cells (9)(10)(11)(12). Other cancer-specific tumor proteins in CML may include mutant p53 and mutant N-ras (12).…”

mentioning

confidence: 99%

Calcium signaling induces acquisition of dendritic cell characteristics in chronic myelogenous leukemia myeloid progenitor cells

Engels

Koski

Bedrosian

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Effective host T lymphocyte sensitization to malignant cells depends on successful antigen presentation. In this study, we examined the capacity of malignant myeloid progenitor cells of patients in the chronic phase of chronic myelogenous leukemia (CML) to acquire characteristics of activated dendritic cells ( Recent clinical observations indicate that T lymphocytes can play an important therapeutic role in chronic myelogenous leukemia (CML) (1, 2). Donor lymphocyte infusions are effective in inducing complete cytogenetic remission in patients who have relapsed after allogeneic bone marrow transplantation, indicating that donor lymphocytes mediate a graftvs.-leukemia response in these patients (3-5).A successful MHC-restricted response against CML is believed to depend on sufficient activation of CD8 ϩ and CD4 ϩ

show abstract

“…The bcr–abl fusion protein can induce specific antibodies (van Denderen et al , 1993) and reactive T cells (Cheever et al , 1993; Fuchs et al , 1995; Komatsu & Moriyama, 1996; Choudhury et al , 1997), and thus the level of bcr–abl transcripts (Miyamura et al , 1994; Hochhaus et al , 1995) and immunogenic proteins may influence the balance between initial growth and survival advantage (Kabarowski et al , 1994) and cell death in CML. Monitoring the transcriptional activity of bcr–abl in individual cells using in situ RT‐PCR in CML may thus also contribute to a better understanding of the natural history of CML.…”

Section: Discussionmentioning

confidence: 99%

Expression of bcr–abl mRNA in individual chronic myelogenous leukaemia cells as determined by in situ amplification

Pachmann¹,

Zhao²,

Schenk³

et al. 2001

Br J Haematol

View full text Add to dashboard Cite

Summary. We present the results of a novel method developed for evaluation of in situ amplification, a molecular genetic method at the cellular level. Reverse transcription polymerase chain reaction (RT-PCR) was used to study bcr± abl transcript levels in individual cells from patients with chronic myelogenous leukaemia (CML). After hybridizing a fluorochrome-labelled probe to the cell-bound RT-PCR product, bcr±abl mRNA-positive cells were determined using image analysis. A dilution series of bcr±abl-positive BV173 into normal cells showed a good correlation between expected and actual values. In 25 CML samples, the percentage of in situ PCR-positive cells showed an excellent correlation with cytogenetic results (r 0´94, P , 0´0001), interphase fluorescence in situ hybridization (FISH) (r 0´95, P 0´001) and hypermetaphase FISH (r 0´81, P , 0´001). The fluorescence intensity was higher in residual CML cells after interferon (IFN) treatment than in newly diagnosed patients (P 0´004), and was highest in late-stage CML resistant to IFN therapy and lowest in CML blast crisis (P 0´001). Mean fluorescence values correlated with bcr±abl protein levels, as determined by Western blot analysis (r 0´62). Laser scanning cytometry allowing automated analysis of large numbers of cells confirmed the results. Thus, fluorescence in situ PCR provides a novel and quantitative approach for monitoring tumour load and bcr± abl transcript levels in CML.

show abstract

Detection of Tumor-Specific Antigens in Philadelphia Chromosome Positive Leukemias

Cited by 8 publications

References 15 publications

Flow cytometric immunobead assay for the detection of BCR–ABL fusion proteins in leukemia patients

Flow cytometric immunobead assay for the detection of BCR–ABL fusion proteins in leukemia patients

Calcium signaling induces acquisition of dendritic cell characteristics in chronic myelogenous leukemia myeloid progenitor cells

Expression of bcr–abl mRNA in individual chronic myelogenous leukaemia cells as determined by in situ amplification

Contact Info

Product

Resources

About