IS CD200 EXPRESSION A SIGNIFICANT MARKER FOR DIFFERENTIATION OF B- LYMPHOPROLIFERATIVE DISEASES? IN A SINGLE TUBE ANALYSIS
(Abstract release date: 05/19/16)
EHA Library. Aytan P. 06/09/16; 134678; PB1778
Dr. Pelin Aytan
Contributions
Contributions
Abstract
Abstract: PB1778
Type: Publication Only
Background
B cells have specific antigen patterns and flow cytometric techniques have been used for discrimination of chronic lymphoproliferative disorders (CLDs). Despite CD23 is considered a reliable marker, because of its positivity in chronic lymphocytic leukemia (CLL) and negativity in mantle cell lymphoma (MCL), there are some cases where CD23 does not discriminate. CD 200 is a trans-membrane glycoprotein belonging to the immunoglobulin super-family mostly expressed by B cells.
Aims
Our aim was to evaluate CD200 expression in the diagnosis and classifications of B-cell CLDs in a single tube by using flow cytometric techniques.
Methods
This is a retrospective study utilizing data on consecutive patients with B CLDs in a single center. The study was performed on erythrocyte-lysed whole peripheral blood samples, after staining with directly conjugated monoclonal antibodies. A single tube was designed to include CD45, CD19, CD5, CD23, CD22, CD10, CD20 and CD200. Samples were analyzed using 8-color MFC (FACS Canto II; BD, Bioscience), and FACS DIVA was used for data analysis. An antigen was considered positive when at least 20% of the cells expressed that antigen.
Results
A total of 59 B-CLD patients - 42 male (71.2%) and 17 female (28.8%) - were evaluated between January 2014 and January 2016. All patients expressed CD19 and while CD20 was found to be positive in all but one CLL case. 36 patients were diagnosed as CLL and 4 patients diagnosed as atypical CLL (aCLL). A total of 19 patients were diagnosed as lymphoma; 3 of them were mantle cell lymphoma (MCL), 2 of them were marginal zone lymphoma (MCL), 2 of them were follicular lymphoma (FL), 2 of them were diffuse large B cell lymphoma (DLBCL) and 2 of them were small lymphocytic lymphoma (SLL). Also 8 patients were unclassified and they were defined as other B lymphoproliferative diseases. While CD200 was positive in all CLL and aCLL patients, it was negative in MCL, MZL, DLBCL patients. CD200 was also positive in unclassified form of B lymphoproliferative diseases in 5 patients. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CD200 for diagnosis of B-CLL are 100%, 52.2%, 76.6% and 100% respectively. These values were 100%, 47.8%, 75% and 100% respectively for CD5. CD 23 was positive in 32 of 36 B-CLL patients (88.9%), and weak positive in the remaining 4 B-CLL patients (11.1%). CD 23 was also found to be positive in 1 (4.3%), weak positive in 4 (17.4%) and negative in 18 (78.3%) patients without B-CLL.
Conclusion
Multiple reports have showed that CD 200 expression is moderate –high in CLL. However, it is dim to negative in mantle cell lymphoma and proposed as another marker to distinguish between CLL and MCL. CLL/ SLL demonstrates low intensity staining for surface immunoglobulin, low expression of CD 22, CD 79B, strong expression of CD 5 and CD 23 but leukemic phase of MCL can be misdiagnosed as CLL. Since MCL is a more aggressive disease and generally treated differently than B-CLL, it is important to differentiate them. In our study, it is showed CD200 expression has a great impact on accurate B-CLDs at diagnosis and it could be added to the routine panels and might be able to show in a single tube by using 8 colour flow cytometer . Also there is high expression of CD200 in B-CLL and it could be a good option for targeted immune therapy in the near future.
Session topic: E-poster
Keyword(s): B cell, CD5, Chronic lymphocytic leukemia
Type: Publication Only
Background
B cells have specific antigen patterns and flow cytometric techniques have been used for discrimination of chronic lymphoproliferative disorders (CLDs). Despite CD23 is considered a reliable marker, because of its positivity in chronic lymphocytic leukemia (CLL) and negativity in mantle cell lymphoma (MCL), there are some cases where CD23 does not discriminate. CD 200 is a trans-membrane glycoprotein belonging to the immunoglobulin super-family mostly expressed by B cells.
Aims
Our aim was to evaluate CD200 expression in the diagnosis and classifications of B-cell CLDs in a single tube by using flow cytometric techniques.
Methods
This is a retrospective study utilizing data on consecutive patients with B CLDs in a single center. The study was performed on erythrocyte-lysed whole peripheral blood samples, after staining with directly conjugated monoclonal antibodies. A single tube was designed to include CD45, CD19, CD5, CD23, CD22, CD10, CD20 and CD200. Samples were analyzed using 8-color MFC (FACS Canto II; BD, Bioscience), and FACS DIVA was used for data analysis. An antigen was considered positive when at least 20% of the cells expressed that antigen.
Results
A total of 59 B-CLD patients - 42 male (71.2%) and 17 female (28.8%) - were evaluated between January 2014 and January 2016. All patients expressed CD19 and while CD20 was found to be positive in all but one CLL case. 36 patients were diagnosed as CLL and 4 patients diagnosed as atypical CLL (aCLL). A total of 19 patients were diagnosed as lymphoma; 3 of them were mantle cell lymphoma (MCL), 2 of them were marginal zone lymphoma (MCL), 2 of them were follicular lymphoma (FL), 2 of them were diffuse large B cell lymphoma (DLBCL) and 2 of them were small lymphocytic lymphoma (SLL). Also 8 patients were unclassified and they were defined as other B lymphoproliferative diseases. While CD200 was positive in all CLL and aCLL patients, it was negative in MCL, MZL, DLBCL patients. CD200 was also positive in unclassified form of B lymphoproliferative diseases in 5 patients. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CD200 for diagnosis of B-CLL are 100%, 52.2%, 76.6% and 100% respectively. These values were 100%, 47.8%, 75% and 100% respectively for CD5. CD 23 was positive in 32 of 36 B-CLL patients (88.9%), and weak positive in the remaining 4 B-CLL patients (11.1%). CD 23 was also found to be positive in 1 (4.3%), weak positive in 4 (17.4%) and negative in 18 (78.3%) patients without B-CLL.
Conclusion
Multiple reports have showed that CD 200 expression is moderate –high in CLL. However, it is dim to negative in mantle cell lymphoma and proposed as another marker to distinguish between CLL and MCL. CLL/ SLL demonstrates low intensity staining for surface immunoglobulin, low expression of CD 22, CD 79B, strong expression of CD 5 and CD 23 but leukemic phase of MCL can be misdiagnosed as CLL. Since MCL is a more aggressive disease and generally treated differently than B-CLL, it is important to differentiate them. In our study, it is showed CD200 expression has a great impact on accurate B-CLDs at diagnosis and it could be added to the routine panels and might be able to show in a single tube by using 8 colour flow cytometer . Also there is high expression of CD200 in B-CLL and it could be a good option for targeted immune therapy in the near future.
Session topic: E-poster
Keyword(s): B cell, CD5, Chronic lymphocytic leukemia
Abstract: PB1778
Type: Publication Only
Background
B cells have specific antigen patterns and flow cytometric techniques have been used for discrimination of chronic lymphoproliferative disorders (CLDs). Despite CD23 is considered a reliable marker, because of its positivity in chronic lymphocytic leukemia (CLL) and negativity in mantle cell lymphoma (MCL), there are some cases where CD23 does not discriminate. CD 200 is a trans-membrane glycoprotein belonging to the immunoglobulin super-family mostly expressed by B cells.
Aims
Our aim was to evaluate CD200 expression in the diagnosis and classifications of B-cell CLDs in a single tube by using flow cytometric techniques.
Methods
This is a retrospective study utilizing data on consecutive patients with B CLDs in a single center. The study was performed on erythrocyte-lysed whole peripheral blood samples, after staining with directly conjugated monoclonal antibodies. A single tube was designed to include CD45, CD19, CD5, CD23, CD22, CD10, CD20 and CD200. Samples were analyzed using 8-color MFC (FACS Canto II; BD, Bioscience), and FACS DIVA was used for data analysis. An antigen was considered positive when at least 20% of the cells expressed that antigen.
Results
A total of 59 B-CLD patients - 42 male (71.2%) and 17 female (28.8%) - were evaluated between January 2014 and January 2016. All patients expressed CD19 and while CD20 was found to be positive in all but one CLL case. 36 patients were diagnosed as CLL and 4 patients diagnosed as atypical CLL (aCLL). A total of 19 patients were diagnosed as lymphoma; 3 of them were mantle cell lymphoma (MCL), 2 of them were marginal zone lymphoma (MCL), 2 of them were follicular lymphoma (FL), 2 of them were diffuse large B cell lymphoma (DLBCL) and 2 of them were small lymphocytic lymphoma (SLL). Also 8 patients were unclassified and they were defined as other B lymphoproliferative diseases. While CD200 was positive in all CLL and aCLL patients, it was negative in MCL, MZL, DLBCL patients. CD200 was also positive in unclassified form of B lymphoproliferative diseases in 5 patients. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CD200 for diagnosis of B-CLL are 100%, 52.2%, 76.6% and 100% respectively. These values were 100%, 47.8%, 75% and 100% respectively for CD5. CD 23 was positive in 32 of 36 B-CLL patients (88.9%), and weak positive in the remaining 4 B-CLL patients (11.1%). CD 23 was also found to be positive in 1 (4.3%), weak positive in 4 (17.4%) and negative in 18 (78.3%) patients without B-CLL.
Conclusion
Multiple reports have showed that CD 200 expression is moderate –high in CLL. However, it is dim to negative in mantle cell lymphoma and proposed as another marker to distinguish between CLL and MCL. CLL/ SLL demonstrates low intensity staining for surface immunoglobulin, low expression of CD 22, CD 79B, strong expression of CD 5 and CD 23 but leukemic phase of MCL can be misdiagnosed as CLL. Since MCL is a more aggressive disease and generally treated differently than B-CLL, it is important to differentiate them. In our study, it is showed CD200 expression has a great impact on accurate B-CLDs at diagnosis and it could be added to the routine panels and might be able to show in a single tube by using 8 colour flow cytometer . Also there is high expression of CD200 in B-CLL and it could be a good option for targeted immune therapy in the near future.
Session topic: E-poster
Keyword(s): B cell, CD5, Chronic lymphocytic leukemia
Type: Publication Only
Background
B cells have specific antigen patterns and flow cytometric techniques have been used for discrimination of chronic lymphoproliferative disorders (CLDs). Despite CD23 is considered a reliable marker, because of its positivity in chronic lymphocytic leukemia (CLL) and negativity in mantle cell lymphoma (MCL), there are some cases where CD23 does not discriminate. CD 200 is a trans-membrane glycoprotein belonging to the immunoglobulin super-family mostly expressed by B cells.
Aims
Our aim was to evaluate CD200 expression in the diagnosis and classifications of B-cell CLDs in a single tube by using flow cytometric techniques.
Methods
This is a retrospective study utilizing data on consecutive patients with B CLDs in a single center. The study was performed on erythrocyte-lysed whole peripheral blood samples, after staining with directly conjugated monoclonal antibodies. A single tube was designed to include CD45, CD19, CD5, CD23, CD22, CD10, CD20 and CD200. Samples were analyzed using 8-color MFC (FACS Canto II; BD, Bioscience), and FACS DIVA was used for data analysis. An antigen was considered positive when at least 20% of the cells expressed that antigen.
Results
A total of 59 B-CLD patients - 42 male (71.2%) and 17 female (28.8%) - were evaluated between January 2014 and January 2016. All patients expressed CD19 and while CD20 was found to be positive in all but one CLL case. 36 patients were diagnosed as CLL and 4 patients diagnosed as atypical CLL (aCLL). A total of 19 patients were diagnosed as lymphoma; 3 of them were mantle cell lymphoma (MCL), 2 of them were marginal zone lymphoma (MCL), 2 of them were follicular lymphoma (FL), 2 of them were diffuse large B cell lymphoma (DLBCL) and 2 of them were small lymphocytic lymphoma (SLL). Also 8 patients were unclassified and they were defined as other B lymphoproliferative diseases. While CD200 was positive in all CLL and aCLL patients, it was negative in MCL, MZL, DLBCL patients. CD200 was also positive in unclassified form of B lymphoproliferative diseases in 5 patients. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of CD200 for diagnosis of B-CLL are 100%, 52.2%, 76.6% and 100% respectively. These values were 100%, 47.8%, 75% and 100% respectively for CD5. CD 23 was positive in 32 of 36 B-CLL patients (88.9%), and weak positive in the remaining 4 B-CLL patients (11.1%). CD 23 was also found to be positive in 1 (4.3%), weak positive in 4 (17.4%) and negative in 18 (78.3%) patients without B-CLL.
Conclusion
Multiple reports have showed that CD 200 expression is moderate –high in CLL. However, it is dim to negative in mantle cell lymphoma and proposed as another marker to distinguish between CLL and MCL. CLL/ SLL demonstrates low intensity staining for surface immunoglobulin, low expression of CD 22, CD 79B, strong expression of CD 5 and CD 23 but leukemic phase of MCL can be misdiagnosed as CLL. Since MCL is a more aggressive disease and generally treated differently than B-CLL, it is important to differentiate them. In our study, it is showed CD200 expression has a great impact on accurate B-CLDs at diagnosis and it could be added to the routine panels and might be able to show in a single tube by using 8 colour flow cytometer . Also there is high expression of CD200 in B-CLL and it could be a good option for targeted immune therapy in the near future.
Session topic: E-poster
Keyword(s): B cell, CD5, Chronic lymphocytic leukemia
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