IMPROTANCE CD138 IMMUNOSTAINING FOR ASSESSMENT OF BONE MARROW PLASMA CELL INFILTRATION IN PATIENTS WITH MULTIPLE MYELOMA
(Abstract release date: 05/19/16)
EHA Library. Usui Y. 06/09/16; 132848; E1299
Dr. Yoshiaki Usui
Contributions
Contributions
Abstract
Abstract: E1299
Type: Eposter Presentation
Background
Estimation of plasma cell (PC) infiltrates in bone marrow (BM) is integral to the diagnosis and monitoring of patients with multiple myeloma (MM). Traditionally, quantification of BM PCs has been performed by differential counting of May–Giemsa-stained aspirate smears. Although previous studies consistently showed a higher % of PCs in BM biopsy sections compared to BM aspirate smears, BM biopsy with CD138 immunostaining has not been widely adopted in clinical practice.
Aims
This study was performed to clarify the importance of CD138 immnostainig for quantification of BM PC by comparing the different methods [BM aspirate clot, BM aspirate smear, and multicolor flow cytometry (FCM)] in patients with MM and monoclonal gammmopathy of undetermined significance (MGUS). In addition, to clarify the clinical relevance of quantifying BM PCs by CD138 immunostaining, 116 patients with monoclonal gammopathy who did not fulfill the criteria of smoldering MM (sMM) based on aspirate smears were reexamined for its diagnosis by BM clots stained with anti-CD138 monoclonal antibody.
Methods
A total of 150 samples of BM biopsy and smears from 100 patients with MGUS and MM, at diagnosis or following therapy were selected. Percentages of PC of aspirated marrow was quantified by May–Giemsa staining, multicolor flowcytometry (MFC), and CD138 immunostianing of BM clot section and biopsy section. PC identification by FCM required at least two markers, i.e., CD38 and either CD45 or CD138. The percentages of PC were calculated relative to the total nucleated cell population. We also investigated the clinical relevance of CD138 immunostaining in the differentiation of MGUS and MM. BM slide sets from 116 untreated patients with M-protein level < 3000 mg/dL in serum or 500 mg/day in urine and without organ damage were re-assessed with based on the % of BM PCs by CD138 immunostaining.
Results
Median % of PC measured by BM biopsy, BM clot, BM smear, and FCM were 13.3%, 12.8 %, 3.7 %, and 2.4 %, respectively. There were significant correlations between BM biopsy and the three other measurement methods, and BM clot showed the strongest correlation with BM biopsy (r = 0.94). In correlation analysis, BM biopsy and BM clot showed significant and very strong correlations in both NCC ≥ 40×103/μL and NCC < 40×103/μL (r = 0.95 and r = 0.91, respectively). Bland–Altman plots showed that BM biopsy agreed well with BM clot but not with BM smear or FCM. There was a significant fixed bias between BM biopsy vs BM smear and BM FCM with mean difference of 15.0% and 16.4%, respectively, indicating that both of these measurements significantly underestimated PCs compared to BM biopsy. However, no significant fixed bias between BM biopsy and BM clot was observed (mean difference: 1.1%). Proportional bias by regressing the differences in values on means of values with linear regression analysis showed significant proportional bias between BM biopsy and BM smear and FCM, but not between BM biopsy and BM clot. As CD138 immunostaining consistently yielded a higher percentage of BM PCs than aspirate smear, it is possible that a considerable portion of patients diagnosed as MGUS by aspirate smear alone could be reclassified as sMM by CD138 immunostaining of BM clot or biopsy. We retrospectively reanalyzed the 116 consecutive patients with monoclonal gammopathy and < 3000 mg/dL or 500 mg/dL M-protein in serum or urine at our hospital. Among the 116 patients that met the above criteria, 13 patients (11%) were classified as sMM as PCs exceeded 10% on aspirate smears. Among the remaining 103 patients, 59 patients (51%) showed > 10% PCs by CD138 immunostaining and was reclassified as sMM and the remaining 44 patients (38%) were still classified as MGUS.
Conclusion
Our data showed that considerable portion of sMM patients could be misclassified as MGUS by bone marrow smear alone and highlight the necessity of CD138 immunostaining of BM biopsy/clot specimens for correct diagnosis.
Session topic: E-poster
Keyword(s): Multiple myeloma, Plasma cells
Type: Eposter Presentation
Background
Estimation of plasma cell (PC) infiltrates in bone marrow (BM) is integral to the diagnosis and monitoring of patients with multiple myeloma (MM). Traditionally, quantification of BM PCs has been performed by differential counting of May–Giemsa-stained aspirate smears. Although previous studies consistently showed a higher % of PCs in BM biopsy sections compared to BM aspirate smears, BM biopsy with CD138 immunostaining has not been widely adopted in clinical practice.
Aims
This study was performed to clarify the importance of CD138 immnostainig for quantification of BM PC by comparing the different methods [BM aspirate clot, BM aspirate smear, and multicolor flow cytometry (FCM)] in patients with MM and monoclonal gammmopathy of undetermined significance (MGUS). In addition, to clarify the clinical relevance of quantifying BM PCs by CD138 immunostaining, 116 patients with monoclonal gammopathy who did not fulfill the criteria of smoldering MM (sMM) based on aspirate smears were reexamined for its diagnosis by BM clots stained with anti-CD138 monoclonal antibody.
Methods
A total of 150 samples of BM biopsy and smears from 100 patients with MGUS and MM, at diagnosis or following therapy were selected. Percentages of PC of aspirated marrow was quantified by May–Giemsa staining, multicolor flowcytometry (MFC), and CD138 immunostianing of BM clot section and biopsy section. PC identification by FCM required at least two markers, i.e., CD38 and either CD45 or CD138. The percentages of PC were calculated relative to the total nucleated cell population. We also investigated the clinical relevance of CD138 immunostaining in the differentiation of MGUS and MM. BM slide sets from 116 untreated patients with M-protein level < 3000 mg/dL in serum or 500 mg/day in urine and without organ damage were re-assessed with based on the % of BM PCs by CD138 immunostaining.
Results
Median % of PC measured by BM biopsy, BM clot, BM smear, and FCM were 13.3%, 12.8 %, 3.7 %, and 2.4 %, respectively. There were significant correlations between BM biopsy and the three other measurement methods, and BM clot showed the strongest correlation with BM biopsy (r = 0.94). In correlation analysis, BM biopsy and BM clot showed significant and very strong correlations in both NCC ≥ 40×103/μL and NCC < 40×103/μL (r = 0.95 and r = 0.91, respectively). Bland–Altman plots showed that BM biopsy agreed well with BM clot but not with BM smear or FCM. There was a significant fixed bias between BM biopsy vs BM smear and BM FCM with mean difference of 15.0% and 16.4%, respectively, indicating that both of these measurements significantly underestimated PCs compared to BM biopsy. However, no significant fixed bias between BM biopsy and BM clot was observed (mean difference: 1.1%). Proportional bias by regressing the differences in values on means of values with linear regression analysis showed significant proportional bias between BM biopsy and BM smear and FCM, but not between BM biopsy and BM clot. As CD138 immunostaining consistently yielded a higher percentage of BM PCs than aspirate smear, it is possible that a considerable portion of patients diagnosed as MGUS by aspirate smear alone could be reclassified as sMM by CD138 immunostaining of BM clot or biopsy. We retrospectively reanalyzed the 116 consecutive patients with monoclonal gammopathy and < 3000 mg/dL or 500 mg/dL M-protein in serum or urine at our hospital. Among the 116 patients that met the above criteria, 13 patients (11%) were classified as sMM as PCs exceeded 10% on aspirate smears. Among the remaining 103 patients, 59 patients (51%) showed > 10% PCs by CD138 immunostaining and was reclassified as sMM and the remaining 44 patients (38%) were still classified as MGUS.
Conclusion
Our data showed that considerable portion of sMM patients could be misclassified as MGUS by bone marrow smear alone and highlight the necessity of CD138 immunostaining of BM biopsy/clot specimens for correct diagnosis.
Session topic: E-poster
Keyword(s): Multiple myeloma, Plasma cells
Abstract: E1299
Type: Eposter Presentation
Background
Estimation of plasma cell (PC) infiltrates in bone marrow (BM) is integral to the diagnosis and monitoring of patients with multiple myeloma (MM). Traditionally, quantification of BM PCs has been performed by differential counting of May–Giemsa-stained aspirate smears. Although previous studies consistently showed a higher % of PCs in BM biopsy sections compared to BM aspirate smears, BM biopsy with CD138 immunostaining has not been widely adopted in clinical practice.
Aims
This study was performed to clarify the importance of CD138 immnostainig for quantification of BM PC by comparing the different methods [BM aspirate clot, BM aspirate smear, and multicolor flow cytometry (FCM)] in patients with MM and monoclonal gammmopathy of undetermined significance (MGUS). In addition, to clarify the clinical relevance of quantifying BM PCs by CD138 immunostaining, 116 patients with monoclonal gammopathy who did not fulfill the criteria of smoldering MM (sMM) based on aspirate smears were reexamined for its diagnosis by BM clots stained with anti-CD138 monoclonal antibody.
Methods
A total of 150 samples of BM biopsy and smears from 100 patients with MGUS and MM, at diagnosis or following therapy were selected. Percentages of PC of aspirated marrow was quantified by May–Giemsa staining, multicolor flowcytometry (MFC), and CD138 immunostianing of BM clot section and biopsy section. PC identification by FCM required at least two markers, i.e., CD38 and either CD45 or CD138. The percentages of PC were calculated relative to the total nucleated cell population. We also investigated the clinical relevance of CD138 immunostaining in the differentiation of MGUS and MM. BM slide sets from 116 untreated patients with M-protein level < 3000 mg/dL in serum or 500 mg/day in urine and without organ damage were re-assessed with based on the % of BM PCs by CD138 immunostaining.
Results
Median % of PC measured by BM biopsy, BM clot, BM smear, and FCM were 13.3%, 12.8 %, 3.7 %, and 2.4 %, respectively. There were significant correlations between BM biopsy and the three other measurement methods, and BM clot showed the strongest correlation with BM biopsy (r = 0.94). In correlation analysis, BM biopsy and BM clot showed significant and very strong correlations in both NCC ≥ 40×103/μL and NCC < 40×103/μL (r = 0.95 and r = 0.91, respectively). Bland–Altman plots showed that BM biopsy agreed well with BM clot but not with BM smear or FCM. There was a significant fixed bias between BM biopsy vs BM smear and BM FCM with mean difference of 15.0% and 16.4%, respectively, indicating that both of these measurements significantly underestimated PCs compared to BM biopsy. However, no significant fixed bias between BM biopsy and BM clot was observed (mean difference: 1.1%). Proportional bias by regressing the differences in values on means of values with linear regression analysis showed significant proportional bias between BM biopsy and BM smear and FCM, but not between BM biopsy and BM clot. As CD138 immunostaining consistently yielded a higher percentage of BM PCs than aspirate smear, it is possible that a considerable portion of patients diagnosed as MGUS by aspirate smear alone could be reclassified as sMM by CD138 immunostaining of BM clot or biopsy. We retrospectively reanalyzed the 116 consecutive patients with monoclonal gammopathy and < 3000 mg/dL or 500 mg/dL M-protein in serum or urine at our hospital. Among the 116 patients that met the above criteria, 13 patients (11%) were classified as sMM as PCs exceeded 10% on aspirate smears. Among the remaining 103 patients, 59 patients (51%) showed > 10% PCs by CD138 immunostaining and was reclassified as sMM and the remaining 44 patients (38%) were still classified as MGUS.
Conclusion
Our data showed that considerable portion of sMM patients could be misclassified as MGUS by bone marrow smear alone and highlight the necessity of CD138 immunostaining of BM biopsy/clot specimens for correct diagnosis.
Session topic: E-poster
Keyword(s): Multiple myeloma, Plasma cells
Type: Eposter Presentation
Background
Estimation of plasma cell (PC) infiltrates in bone marrow (BM) is integral to the diagnosis and monitoring of patients with multiple myeloma (MM). Traditionally, quantification of BM PCs has been performed by differential counting of May–Giemsa-stained aspirate smears. Although previous studies consistently showed a higher % of PCs in BM biopsy sections compared to BM aspirate smears, BM biopsy with CD138 immunostaining has not been widely adopted in clinical practice.
Aims
This study was performed to clarify the importance of CD138 immnostainig for quantification of BM PC by comparing the different methods [BM aspirate clot, BM aspirate smear, and multicolor flow cytometry (FCM)] in patients with MM and monoclonal gammmopathy of undetermined significance (MGUS). In addition, to clarify the clinical relevance of quantifying BM PCs by CD138 immunostaining, 116 patients with monoclonal gammopathy who did not fulfill the criteria of smoldering MM (sMM) based on aspirate smears were reexamined for its diagnosis by BM clots stained with anti-CD138 monoclonal antibody.
Methods
A total of 150 samples of BM biopsy and smears from 100 patients with MGUS and MM, at diagnosis or following therapy were selected. Percentages of PC of aspirated marrow was quantified by May–Giemsa staining, multicolor flowcytometry (MFC), and CD138 immunostianing of BM clot section and biopsy section. PC identification by FCM required at least two markers, i.e., CD38 and either CD45 or CD138. The percentages of PC were calculated relative to the total nucleated cell population. We also investigated the clinical relevance of CD138 immunostaining in the differentiation of MGUS and MM. BM slide sets from 116 untreated patients with M-protein level < 3000 mg/dL in serum or 500 mg/day in urine and without organ damage were re-assessed with based on the % of BM PCs by CD138 immunostaining.
Results
Median % of PC measured by BM biopsy, BM clot, BM smear, and FCM were 13.3%, 12.8 %, 3.7 %, and 2.4 %, respectively. There were significant correlations between BM biopsy and the three other measurement methods, and BM clot showed the strongest correlation with BM biopsy (r = 0.94). In correlation analysis, BM biopsy and BM clot showed significant and very strong correlations in both NCC ≥ 40×103/μL and NCC < 40×103/μL (r = 0.95 and r = 0.91, respectively). Bland–Altman plots showed that BM biopsy agreed well with BM clot but not with BM smear or FCM. There was a significant fixed bias between BM biopsy vs BM smear and BM FCM with mean difference of 15.0% and 16.4%, respectively, indicating that both of these measurements significantly underestimated PCs compared to BM biopsy. However, no significant fixed bias between BM biopsy and BM clot was observed (mean difference: 1.1%). Proportional bias by regressing the differences in values on means of values with linear regression analysis showed significant proportional bias between BM biopsy and BM smear and FCM, but not between BM biopsy and BM clot. As CD138 immunostaining consistently yielded a higher percentage of BM PCs than aspirate smear, it is possible that a considerable portion of patients diagnosed as MGUS by aspirate smear alone could be reclassified as sMM by CD138 immunostaining of BM clot or biopsy. We retrospectively reanalyzed the 116 consecutive patients with monoclonal gammopathy and < 3000 mg/dL or 500 mg/dL M-protein in serum or urine at our hospital. Among the 116 patients that met the above criteria, 13 patients (11%) were classified as sMM as PCs exceeded 10% on aspirate smears. Among the remaining 103 patients, 59 patients (51%) showed > 10% PCs by CD138 immunostaining and was reclassified as sMM and the remaining 44 patients (38%) were still classified as MGUS.
Conclusion
Our data showed that considerable portion of sMM patients could be misclassified as MGUS by bone marrow smear alone and highlight the necessity of CD138 immunostaining of BM biopsy/clot specimens for correct diagnosis.
Session topic: E-poster
Keyword(s): Multiple myeloma, Plasma cells
{{ help_message }}
{{filter}}