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The influence of cytotoxic drugs on the immunophenotype of blast cells in paediatric B precursor acute lymphoblastic leukaemia Cover

The influence of cytotoxic drugs on the immunophenotype of blast cells in paediatric B precursor acute lymphoblastic leukaemia

Radiology and Oncology
Volume 58 (2024): Issue 1 (March 2024)
By:
Open Access
|Feb 2024

Figures & Tables

FIGURE 1.

Example of a gating procedure. Mononuclear cells were selected on the forward versus side scatter plot (A) and single cells by gating on the area against the width for forward scatter (B). Propidium iodide (PI) was used to select viable cells (C). The final selection of blasts was based on their immunophenotype, in this case by selecting CD10-positive cells on the CD10 versus CD45 plots (D).
Example of a gating procedure. Mononuclear cells were selected on the forward versus side scatter plot (A) and single cells by gating on the area against the width for forward scatter (B). Propidium iodide (PI) was used to select viable cells (C). The final selection of blasts was based on their immunophenotype, in this case by selecting CD10-positive cells on the CD10 versus CD45 plots (D).

FIGURE 2A.

The plots of the marginal effects for the fixed effects coefficients of the models for CD137, CD34 and CD19. The lines correspond to the estimates of the ma rginal ef fects and the coloured areas to the 95% confidence intervals.
The plots of the marginal effects for the fixed effects coefficients of the models for CD137, CD34 and CD19. The lines correspond to the estimates of the ma rginal ef fects and the coloured areas to the 95% confidence intervals.

FIGURE 2B.

The plots of the marginal effects for the fixed effects coefficients of the models for CD20, CD45, CD27, CD66c, CD10, and CD58. The lines correspond to the estimates of the marginal effects and the coloured areas to the 95% confidence intervals.
The plots of the marginal effects for the fixed effects coefficients of the models for CD20, CD45, CD27, CD66c, CD10, and CD58. The lines correspond to the estimates of the marginal effects and the coloured areas to the 95% confidence intervals.

Response to the therapy in patients treated according ALL IC BFM 09 protocol

Patients treated according to ALL IC-BFM 2009 protocol
No. of patients27
Treatment armN (%)
  SR6 (22)
  IR18 (66)
  HR3 (11)
Response to prednisolone on day 8N (%)
  good response*27 (100)
  poor response**0 (0)
Minimal residual disease on day 15N (%)
  < 0.1%10 (37)
  0.1–10%15 (56)
  > 10%2 (7)
Minimal residual disease on day 33N (%)
  neg16 (59)
  < 0.1%6 (22)
  0.1–10%3 (11)
  > 10%2 (7)
OutcomeN (%)
  alive26 (96)
  died1 (4)

Clinical characteristics of all included patients

No. of all included patients with B-ALLN = 29
Male/femaleN
17/12
Mean age at diagnosisYears
4.76 (newborn 17)
Flow cytometric findings at diagnosisN (%)
  Pro-B3 (10)
  Pre-B6 (21)
  Common type20 (69)
Cytogenetic and molecular finding at diagnosisN (%)
  normal karyotype6 (21)
  hyperdiploid10 (34)
  t(12;21)(28)
  complex karyotype(3)
  MLL1 (3)
  changes of unknown risk potential3 (10)
OutcomeN (%)
  Alive93)
  Died2 (7)
Cause of deathN
  pancreatitis*1
  progressive disease**1

The description of the antibodies used for flow cytometry

AntibodyAntibody cloneAntibody vendorAntibody volume (μL)
CD10Anti-CD10 Horizon™ BV605HI10aBD Biosciences5
CD19CD19-BD Horizon™ V450 Mouse Anti-Human CD19SJ25C1BD Biosciences5
CD20CD20-BD™ CD20 APCL27BD Biosciences7
CD27CD27-BD Horizon™ APC-R700 Mouse Anti-Human CD27M-T271BD Biosciences5
CD34CD34-BD™ CD34 PE-Cy™78G12BD Biosciences5
CD45CD45-BD™ CD45 APC-Cy™72D1BD Biosciences3
CD58CD58-BD Pharmingen™ FITC Mouse Anti-Human CD581C3BD Biosciences3
CD66cCD66c-BD OptiBuild™ BV510 Mouse Anti-Human CD66cB6.2/CD66BD Biosciences5
CD137CD137-BD Pharmingen™ PE Mouse Anti-Human CD1374B4-1BD Biosciences7

Antigens expressed on acute lymphoblastic leukaemia (ALL) blasts which are essential for planning the treatment or may influence the prognosis of the disease

AntigenExpression of the antigen in normal and pathological conditionsClinical significance
CD10Normal lymphoid progenitors, neutrophils and blast cells of B-ALL19Associated with favourable presenting features in paediatric B-ALL and possible target for emerging CAR-T19,20,21
CD19Normal and malignant B lymphocytes22Target of blinatumomab and CAR-T22,23,24
CD20Normal and malignant B lymphocytes21Target of rituximab, up-modulated in B-ALL patients with poor prognosis7,21
CD27T cells, natural killer cells and thymocytes, also memory B cells, and in some subsets of B-ALL11Positive in B-ALL with BCR/ABL or CRLF2 rearrangement, high positivity was associated with poor prognosis11,12,13
CD34Pluripotent stem cells14Lack of expression associated with worse event free and overall survival of ALL15
CD45Cells of hematopoietic origin25High expression associated with high risk disease and worse event free survival, and worse rate of complete remission after the induction therapy16,26
CD58Hematopoietic and nonhematopoietic cells27Lack of expression reduces the efficacy of blinatumomab and anti-CD19 CAR-T4,5
CD66cGranulocytes and their precursors, most common myeloid antigen on malignant B lymphoblasts17Associated with specific genetic alterations such as BCR/ABL, hypodiploidy, hypodiploidy and CRLF2-positivity in B-ALL17,18
CD137T lymphocytes and natural killer cells but also on activated B cells of naive origin28Enhances B cell survival28

The coefficients obtained by using separate mixed model which showed the effect of single drug in CDn antigen expression_ The coefficient d showed the presence of CDn antigen and coefficient of k_d the influence of drug dose on antigen expression_ The way how the given drug influences the antigen expression is shown in different colours: RED (coefficients d and k_d are statistically significantly different from 0), BLUE (the coefficient d is significantly different from 0), GREEN (coefficient k_d is significantly different from 0) and WHITE (neither the coefficient d nor k_d is statistically significantly different from 0)_ 95% confidence intervals, and corresponding p values are also shown_ All confidence intervals and p values were adjusted with Holm’s method

DrugCDCoefficient d (presence)p value for dCoefficient k_d (log(amount))p value for k_d
AsparaginaseCD10−0.12, [−0.33, 0.1]1−0.01, [−0.11, 0.1]1
CD137−0.18, [−0.46, 0.1]10, [−0.11, 0.12]1
CD19−0.09, [−0.42, 0.24]1−0.01, [−0.14, 0.13]1
CD20−0.15, [−0.39, 0.09]10, [−0.09, 0.1]1
CD27−0.03, [−0.28, 0.22]1−0.01, [−0.12, 0.11]1
CD34−0.15, [−0.4, 0.09]1−0.09, [−0.23, 0.04]1
CD45−0.04, [−0.27, 0.2]1−0.02, [−0.16, 0.11]1
CD58−0.05, [−0.23, 0.13]10.01, [−0.08, 0.1]1
CD66c−0.05, [−0.36, 0.27]10.02, [−0.15, 0.19]1
DaunorubicinCD10−1.1, [−1.38, −0.83]< 0.001−0.47, [−0.63, −0.31]< 0.001
CD1371.05, [0.72, 1.37]< 0.0010.4, [0.21, 0.59]< 0.001
CD19−1.05, [−1.48, −0.62]< 0.001−0.35, [−0.6, −0.1]< 0.001
CD20−0.24, [−0.53, 0.05]0.35−0.22, [−0.39, −0.05]0.002
CD27−0.6, [−0.92, −0.27]< 0.001−0.09, [−0.27, 0.09]1
CD34−1.48, [−1.8, −1.16]< 0.001−0.74, [−0.93, −0.55]< 0.001
CD45−0.58, [−0.9, −0.27]< 0.001−0.28, [−0.47, −0.1]< 0.001
CD58−0.73, [−0.97, −0.5]< 0.001−0.32, [−0.46, −0.19]< 0.001
CD66c0.28, [−0.13, 0.69]10.13, [−0.1, 0.37]1
MethotrexateCD10−0.1, [−0.52, 0.33]1−0.04, [−0.32, 0.24]1
CD1370.11, [−0.33, 0.56]10.02, [−0.24, 0.29]1
CD19−0.33, [−1, 0.35]10.02, [−0.38, 0.42]1
CD20−0.21, [−0.68, 0.27]10.04, [−0.26, 0.34]1
CD27−0.26, [−0.79, 0.27]1−0.03, [−0.34, 0.29]1
CD340.08, [−0.4, 0.56]10.04, [−0.28, 0.35]1
CD45−0.04, [−0.5, 0.41]10.03, [−0.29, 0.35]1
CD580.01, [−0.25, 0.27]10.02, [−0.21, 0.26]1
CD66c−0.05, [−0.64, 0.54]1−0.02, [−0.4, 0.37]1
PrednisoloneCD10−0.42, [−0.64, −0.21]< 0.001−0.01, [−0.12, 0.1]1
CD137−0.06, [−0.32, 0.19]10.05, [−0.1, 0.19]1
CD19−0.61, [−0.94, −0.27]< 0.001−0.03, [−0.2, 0.15]1
CD200.14, [−0.09, 0.37]10.02, [−0.1, 0.14]1
CD27−0.5, [−0.76, −0.23]< 0.001−0.15, [−0.3, 0]0.055
CD34−0.35, [−0.6, −0.11]< 0.0010.03, [−0.1, 0.16]1
CD45−0.12, [−0.36, 0.12]1−0.07, [−0.2, 0.07]1
CD58−0.19, [−0.38, −0.01]0.034−0.05, [−0.15, 0.05]1
CD66c0, [−0.19, 0.19]1−0.02, [−0.2, 0.15]1
VincristineCD10−0.14, [−0.37, 0.1]1−0.05, [−0.14, 0.05]1
CD1370.01, [−0.22, 0.25]10.07, [−0.04, 0.18]1
CD19−0.56, [−0.94, 0]< 0.001−0.17, [−0.32, −0.03]0.004
CD200.06, [−0.19, 0.31]10.01, [−0.08, 0.09]1
CD27−0.06, [−0.34, 0.23]10, [−0.09, 0.09]1
CD34−0.18, [−0.45, 0.1]1−0.04, [−0.14, 0.06]1
CD450.36, [0.07, 0.64]0.0020.16, [0.05, 0.26]< 0.001
D58−0.27, [−0.48, −0.06]0.001−0.11, [−0.19, −0.03]< 0.001
CD66c−0.04, [−0.39, 0.3]1−0.02, [−0.15, 0.11]1
DOI: https://doi.org/10.2478/raon-2024-0006 | Journal eISSN: 1581-3207 | Journal ISSN: 1318-2099
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Language: English
Page range: 133 - 144
Submitted on: May 14, 2023
Accepted on: Dec 6, 2023
Published on: Feb 21, 2024
Published by: Association of Radiology and Oncology
In partnership with: Paradigm Publishing Services
Publication frequency: 4 times per year
Keywords:
immunophenotypic changes,
chemotherapy,
immunotherapy,
paediatric B-ALL
Related subjects:
Medicine,
Clinical medicine,
Internal medicine,
Haematology, oncology,
Radiology

© 2024 Tomaz Prelog, Simon Bucek, Andreja Brozic, Jakob Peterlin, Marko Kavcic, Masa Omerzel, Bostjan Markelc, Tanja Jesenko, Veronika Kloboves Prevodnik, published by Association of Radiology and Oncology
This work is licensed under the Creative Commons Attribution 4.0 License.

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