FLOW CYTOMETRY OF CEREBROSPINAL FLUID IN CHILDREN WITH ACUTE LYMPHOBLASTIC LEUKEMIA AT DIAGNOSIS AND RELAPSE
(Abstract release date: 05/19/16)
EHA Library. Gabelli M. 06/09/16; 132410; E861
Dr. Maria Gabelli
Contributions
Contributions
Abstract
Abstract: E861
Type: Eposter Presentation
Background
Acute lymphoblastic leukemia (ALL) may involve central nervous system (CNS) in 3-6% of pediatric patients. Cytology (CC) of cerebrospinal fluid (CSF) is currently used to define CNS infiltration, although sensitivity and specificity are low. Flow cytometry (FC) can identify blasts in CSF samples that are negative for cytology with higher sensitivity and specificity. The frequency and the significance of this occult cerebrospinal involvement in children with ALL is still not clear.
Aims
We explored the feasibility of CSF flow cytometric analysis at each lumbar puncture during therapy in primary and relapsed ALL. Moreover, we studied prospectively its clinical significance in comparison with cytology and cell count.
Methods
We included children (aged 1-18 years) with Philadelphia negative ALL and with ALL isolated marrow relapse diagnosed from 12.09.2013 to 31.01.2016 at our Institution, after informed consent acquisition. Treatment schedule and definition of CSF involvement were as per AIEOP-BFM ALL2009 Protocol. At each point of intrathecal therapy, CSF was collected and analyzed within 24 hours by an automated cell counter, by cytology and 8-color flow cytometry (B or T lineage panel). A tiny cluster of events with immunophenotype compatible with blasts at diagnosis was considered positive by FC (FC+).
Results
784 samples from 76 patients with de novo ALL and 92 samples from 13 children with marrow relapse were considered. Median follow up was 14 months (1-28,5 months). At diagnosis 4 T-ALL patients (5%) presented CNS involvement (CNS3): 3 with positive cytology (CC+) and positive FC, 1 with cranial nerve palsy (CC-/FC-). Other 3 patients (pB-ALL) had positive cytology but <5 cell/µl (CNS2): 2 were FC+, 1 FC- (normal T and B cells). 25/76 children (33%) with no CNS leukemia (CNS1) resulted FC+: 3 T-ALL, 22 pB-ALL. 9/25 (36%) CNS1/FC+ patients were high risk (MRD based stratification). FC persisted positive at day 15 in 2 CNS3/FC+ patients, in 1 CNS2/FC+ and in 2 patients that were positive only by FC at diagnosis. Among patients who were FC+ at diagnosis, we found other isolated FC+ samples at subsequent time points: 2 during induction, 4 during consolidation. Moreover, during reinduction a child CNS3/FC+ at diagnosis presented other 3 samples FC+/CC-. Even children who had negative CSF by flow cytometry at diagnosis presented isolated FC+ samples at later time points: 1 during induction, 6 during consolidation. One child (CNS1/FC- at diagnosis) died for infection, one patient (CNS1/FC+ at diagnosis) presented early marrow relapse, the remaining 74 are alive in complete remission (4/74 after hematopoietic stem cell transplantation). Among 13 patients with isolated marrow ALL relapse, 7 (54%) had FC+ CSF at presentation. 4/7 had a second relapse (2 CNS relapse, 2 BM relapse), 3 died. Among 6 FC- patients, 1 had a second marrow relapse, 1 died of transplant related mortality, 1 showed disease progression and 3 are alive and in complete remission.
Conclusion
Flow Cytometry of CSF is a simple, rapid and cheap method that is able to identify blasts with high sensibility and specificity, allowing the detection of occult CNS infiltration. Our preliminary data suggest that FC CSF positivity may be a risk factor for BM and CNS relapse in children. A longer follow up and a larger group of patients are needed to confirm these findings in ALL at diagnosis and relapse.
Session topic: E-poster
Keyword(s): Acute lymphoblastic leukemia, Childhood, Flow cytometry
Type: Eposter Presentation
Background
Acute lymphoblastic leukemia (ALL) may involve central nervous system (CNS) in 3-6% of pediatric patients. Cytology (CC) of cerebrospinal fluid (CSF) is currently used to define CNS infiltration, although sensitivity and specificity are low. Flow cytometry (FC) can identify blasts in CSF samples that are negative for cytology with higher sensitivity and specificity. The frequency and the significance of this occult cerebrospinal involvement in children with ALL is still not clear.
Aims
We explored the feasibility of CSF flow cytometric analysis at each lumbar puncture during therapy in primary and relapsed ALL. Moreover, we studied prospectively its clinical significance in comparison with cytology and cell count.
Methods
We included children (aged 1-18 years) with Philadelphia negative ALL and with ALL isolated marrow relapse diagnosed from 12.09.2013 to 31.01.2016 at our Institution, after informed consent acquisition. Treatment schedule and definition of CSF involvement were as per AIEOP-BFM ALL2009 Protocol. At each point of intrathecal therapy, CSF was collected and analyzed within 24 hours by an automated cell counter, by cytology and 8-color flow cytometry (B or T lineage panel). A tiny cluster of events with immunophenotype compatible with blasts at diagnosis was considered positive by FC (FC+).
Results
784 samples from 76 patients with de novo ALL and 92 samples from 13 children with marrow relapse were considered. Median follow up was 14 months (1-28,5 months). At diagnosis 4 T-ALL patients (5%) presented CNS involvement (CNS3): 3 with positive cytology (CC+) and positive FC, 1 with cranial nerve palsy (CC-/FC-). Other 3 patients (pB-ALL) had positive cytology but <5 cell/µl (CNS2): 2 were FC+, 1 FC- (normal T and B cells). 25/76 children (33%) with no CNS leukemia (CNS1) resulted FC+: 3 T-ALL, 22 pB-ALL. 9/25 (36%) CNS1/FC+ patients were high risk (MRD based stratification). FC persisted positive at day 15 in 2 CNS3/FC+ patients, in 1 CNS2/FC+ and in 2 patients that were positive only by FC at diagnosis. Among patients who were FC+ at diagnosis, we found other isolated FC+ samples at subsequent time points: 2 during induction, 4 during consolidation. Moreover, during reinduction a child CNS3/FC+ at diagnosis presented other 3 samples FC+/CC-. Even children who had negative CSF by flow cytometry at diagnosis presented isolated FC+ samples at later time points: 1 during induction, 6 during consolidation. One child (CNS1/FC- at diagnosis) died for infection, one patient (CNS1/FC+ at diagnosis) presented early marrow relapse, the remaining 74 are alive in complete remission (4/74 after hematopoietic stem cell transplantation). Among 13 patients with isolated marrow ALL relapse, 7 (54%) had FC+ CSF at presentation. 4/7 had a second relapse (2 CNS relapse, 2 BM relapse), 3 died. Among 6 FC- patients, 1 had a second marrow relapse, 1 died of transplant related mortality, 1 showed disease progression and 3 are alive and in complete remission.
Conclusion
Flow Cytometry of CSF is a simple, rapid and cheap method that is able to identify blasts with high sensibility and specificity, allowing the detection of occult CNS infiltration. Our preliminary data suggest that FC CSF positivity may be a risk factor for BM and CNS relapse in children. A longer follow up and a larger group of patients are needed to confirm these findings in ALL at diagnosis and relapse.
Session topic: E-poster
Keyword(s): Acute lymphoblastic leukemia, Childhood, Flow cytometry
Abstract: E861
Type: Eposter Presentation
Background
Acute lymphoblastic leukemia (ALL) may involve central nervous system (CNS) in 3-6% of pediatric patients. Cytology (CC) of cerebrospinal fluid (CSF) is currently used to define CNS infiltration, although sensitivity and specificity are low. Flow cytometry (FC) can identify blasts in CSF samples that are negative for cytology with higher sensitivity and specificity. The frequency and the significance of this occult cerebrospinal involvement in children with ALL is still not clear.
Aims
We explored the feasibility of CSF flow cytometric analysis at each lumbar puncture during therapy in primary and relapsed ALL. Moreover, we studied prospectively its clinical significance in comparison with cytology and cell count.
Methods
We included children (aged 1-18 years) with Philadelphia negative ALL and with ALL isolated marrow relapse diagnosed from 12.09.2013 to 31.01.2016 at our Institution, after informed consent acquisition. Treatment schedule and definition of CSF involvement were as per AIEOP-BFM ALL2009 Protocol. At each point of intrathecal therapy, CSF was collected and analyzed within 24 hours by an automated cell counter, by cytology and 8-color flow cytometry (B or T lineage panel). A tiny cluster of events with immunophenotype compatible with blasts at diagnosis was considered positive by FC (FC+).
Results
784 samples from 76 patients with de novo ALL and 92 samples from 13 children with marrow relapse were considered. Median follow up was 14 months (1-28,5 months). At diagnosis 4 T-ALL patients (5%) presented CNS involvement (CNS3): 3 with positive cytology (CC+) and positive FC, 1 with cranial nerve palsy (CC-/FC-). Other 3 patients (pB-ALL) had positive cytology but <5 cell/µl (CNS2): 2 were FC+, 1 FC- (normal T and B cells). 25/76 children (33%) with no CNS leukemia (CNS1) resulted FC+: 3 T-ALL, 22 pB-ALL. 9/25 (36%) CNS1/FC+ patients were high risk (MRD based stratification). FC persisted positive at day 15 in 2 CNS3/FC+ patients, in 1 CNS2/FC+ and in 2 patients that were positive only by FC at diagnosis. Among patients who were FC+ at diagnosis, we found other isolated FC+ samples at subsequent time points: 2 during induction, 4 during consolidation. Moreover, during reinduction a child CNS3/FC+ at diagnosis presented other 3 samples FC+/CC-. Even children who had negative CSF by flow cytometry at diagnosis presented isolated FC+ samples at later time points: 1 during induction, 6 during consolidation. One child (CNS1/FC- at diagnosis) died for infection, one patient (CNS1/FC+ at diagnosis) presented early marrow relapse, the remaining 74 are alive in complete remission (4/74 after hematopoietic stem cell transplantation). Among 13 patients with isolated marrow ALL relapse, 7 (54%) had FC+ CSF at presentation. 4/7 had a second relapse (2 CNS relapse, 2 BM relapse), 3 died. Among 6 FC- patients, 1 had a second marrow relapse, 1 died of transplant related mortality, 1 showed disease progression and 3 are alive and in complete remission.
Conclusion
Flow Cytometry of CSF is a simple, rapid and cheap method that is able to identify blasts with high sensibility and specificity, allowing the detection of occult CNS infiltration. Our preliminary data suggest that FC CSF positivity may be a risk factor for BM and CNS relapse in children. A longer follow up and a larger group of patients are needed to confirm these findings in ALL at diagnosis and relapse.
Session topic: E-poster
Keyword(s): Acute lymphoblastic leukemia, Childhood, Flow cytometry
Type: Eposter Presentation
Background
Acute lymphoblastic leukemia (ALL) may involve central nervous system (CNS) in 3-6% of pediatric patients. Cytology (CC) of cerebrospinal fluid (CSF) is currently used to define CNS infiltration, although sensitivity and specificity are low. Flow cytometry (FC) can identify blasts in CSF samples that are negative for cytology with higher sensitivity and specificity. The frequency and the significance of this occult cerebrospinal involvement in children with ALL is still not clear.
Aims
We explored the feasibility of CSF flow cytometric analysis at each lumbar puncture during therapy in primary and relapsed ALL. Moreover, we studied prospectively its clinical significance in comparison with cytology and cell count.
Methods
We included children (aged 1-18 years) with Philadelphia negative ALL and with ALL isolated marrow relapse diagnosed from 12.09.2013 to 31.01.2016 at our Institution, after informed consent acquisition. Treatment schedule and definition of CSF involvement were as per AIEOP-BFM ALL2009 Protocol. At each point of intrathecal therapy, CSF was collected and analyzed within 24 hours by an automated cell counter, by cytology and 8-color flow cytometry (B or T lineage panel). A tiny cluster of events with immunophenotype compatible with blasts at diagnosis was considered positive by FC (FC+).
Results
784 samples from 76 patients with de novo ALL and 92 samples from 13 children with marrow relapse were considered. Median follow up was 14 months (1-28,5 months). At diagnosis 4 T-ALL patients (5%) presented CNS involvement (CNS3): 3 with positive cytology (CC+) and positive FC, 1 with cranial nerve palsy (CC-/FC-). Other 3 patients (pB-ALL) had positive cytology but <5 cell/µl (CNS2): 2 were FC+, 1 FC- (normal T and B cells). 25/76 children (33%) with no CNS leukemia (CNS1) resulted FC+: 3 T-ALL, 22 pB-ALL. 9/25 (36%) CNS1/FC+ patients were high risk (MRD based stratification). FC persisted positive at day 15 in 2 CNS3/FC+ patients, in 1 CNS2/FC+ and in 2 patients that were positive only by FC at diagnosis. Among patients who were FC+ at diagnosis, we found other isolated FC+ samples at subsequent time points: 2 during induction, 4 during consolidation. Moreover, during reinduction a child CNS3/FC+ at diagnosis presented other 3 samples FC+/CC-. Even children who had negative CSF by flow cytometry at diagnosis presented isolated FC+ samples at later time points: 1 during induction, 6 during consolidation. One child (CNS1/FC- at diagnosis) died for infection, one patient (CNS1/FC+ at diagnosis) presented early marrow relapse, the remaining 74 are alive in complete remission (4/74 after hematopoietic stem cell transplantation). Among 13 patients with isolated marrow ALL relapse, 7 (54%) had FC+ CSF at presentation. 4/7 had a second relapse (2 CNS relapse, 2 BM relapse), 3 died. Among 6 FC- patients, 1 had a second marrow relapse, 1 died of transplant related mortality, 1 showed disease progression and 3 are alive and in complete remission.
Conclusion
Flow Cytometry of CSF is a simple, rapid and cheap method that is able to identify blasts with high sensibility and specificity, allowing the detection of occult CNS infiltration. Our preliminary data suggest that FC CSF positivity may be a risk factor for BM and CNS relapse in children. A longer follow up and a larger group of patients are needed to confirm these findings in ALL at diagnosis and relapse.
Session topic: E-poster
Keyword(s): Acute lymphoblastic leukemia, Childhood, Flow cytometry
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