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Immunophenotyping of Waldenströms macroglobulinemia cell lines reveals distinct patterns of surface antigen expression: potential biological and therapeutic implications.

Paulus A, Chitta KS, Wallace PK, Advani PP, Akhtar S, Kuranz-Blake M, Ailawadhi S, Chanan-Khan AA - PLoS ONE (2015)

Bottom Line: RPCI-WM1 cells demonstrated decreased expression of CD19, CD20, and CD23 with enhanced expression of CD28, CD38 and CD184, antigens that were differentially expressed on BCWM.1 and MWCL-1 cells.Due to increased expression of CD184/CXCR4 and CD38, RPCI-WM1 represents a valuable model in which to study the effects anti-CXCR4 or anti-CD38 targeted therapies that are actively being developed for treatment of hematologic cancers.Our analysis defines the utility of the most commonly employed WM cell lines as based on their immunophenotype profiles, highlighting unique differences that can be further studied for therapeutic exploit.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Mayo Clinic, Jacksonville, Florida, United States of America.

ABSTRACT
Waldenströms macroglobulinemia (WM) is a subtype of Non-Hodgkin's lymphoma in which the tumor cell population is markedly heterogeneous, consisting of immunoglobulin-M secreting B-lymphocytes, plasmacytoid lymphocytes and plasma cells. Due to rarity of disease and scarcity of reliable preclinical models, many facets of WM molecular and phenotypic architecture remain incompletely understood. Currently, there are 3 human WM cell lines that are routinely used in experimental studies, namely, BCWM.1, MWCL-1 and RPCI-WM1. During establishment of RPCI-WM1, we observed loss of the CD19 and CD20 antigens, which are typically present on WM cells. Intrigued by this observation and in an effort to better define the immunophenotypic makeup of this cell line, we conducted a more comprehensive analysis for the presence or absence of other cell surface antigens that are present on the RPCI-WM1 model, as well as those on the two other WM cell lines, BCWM.1 and MWCL-1. We examined expression of 65 extracellular and 4 intracellular antigens, comprising B-cell, plasma cell, T-cell, NK-cell, myeloid and hematopoietic stem cell surface markers by flow cytometry analysis. RPCI-WM1 cells demonstrated decreased expression of CD19, CD20, and CD23 with enhanced expression of CD28, CD38 and CD184, antigens that were differentially expressed on BCWM.1 and MWCL-1 cells. Due to increased expression of CD184/CXCR4 and CD38, RPCI-WM1 represents a valuable model in which to study the effects anti-CXCR4 or anti-CD38 targeted therapies that are actively being developed for treatment of hematologic cancers. Overall, differences in surface antigen expression across the 3 cell lines may reflect the tumor clone population predominant in the index patients, from whom the cell lines were developed. Our analysis defines the utility of the most commonly employed WM cell lines as based on their immunophenotype profiles, highlighting unique differences that can be further studied for therapeutic exploit.

No MeSH data available.


Related in: MedlinePlus

WM-specific antigen expression compared across RPCI-WM1, BCWM.1 and MWCL-1 cell lines.Fluorescein (FITC), phycoerythrin (PE), phycoerythrin—cyanine 5 (PC5) or allophycocyanin (APC) conjugates of various antibodies were used as presented above. All cell lines were negative for CD10 and 11c. Blue line indicates RPCI-WM1 antigen expression, red line indicates MWCL-1 antigen expression and green line indicates antigen expression in BCWM.1 cell line. Only BCWM.1 an MWCL-1 were CD19+, CD20+ and FMC7+. Expression of CD138 and κ-light-chain was seen only on MWCL-1 and RPCI-WM1 cells. CD28 expression was markedly more observable on RPCI-WM1 tumor cells as compared to MWCL-1 or BCWM.1.
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pone.0122338.g002: WM-specific antigen expression compared across RPCI-WM1, BCWM.1 and MWCL-1 cell lines.Fluorescein (FITC), phycoerythrin (PE), phycoerythrin—cyanine 5 (PC5) or allophycocyanin (APC) conjugates of various antibodies were used as presented above. All cell lines were negative for CD10 and 11c. Blue line indicates RPCI-WM1 antigen expression, red line indicates MWCL-1 antigen expression and green line indicates antigen expression in BCWM.1 cell line. Only BCWM.1 an MWCL-1 were CD19+, CD20+ and FMC7+. Expression of CD138 and κ-light-chain was seen only on MWCL-1 and RPCI-WM1 cells. CD28 expression was markedly more observable on RPCI-WM1 tumor cells as compared to MWCL-1 or BCWM.1.

Mentions: We then conducted a comprehensive comparative analysis of surface markers in WM cell lines (only 3 noted in the medical literature, 2 developed by the Mayo Clinic group; MWCL-1 and RPCI-WM1,[10, 14] and 1 developed at Dana Farber Cancer Institute; BCWM.1).[13] Using the same set of 19 antigens (Fig 1) a comparative analysis was performed in BCWM.1 and MWCL-1 (Fig 2). Both MWCL-1 and BCWM.1 cells were CD19+low and CD20+medium. The CD20 epitope, FMC7, was also expressed in ~45–63% of BCWM.1 and MWCL-1 cell, respectively albeit at low levels. Notably both RPCI-WM1 (88.7% of cells) and MWCL-1 (98.3% of cells) were CD138+medium. RPCI-WM1 (99.1% of cells) demonstrated low density of κ light-chain (MESF 10,789.6) whereas density of κ light-chain on MWCL-1 (99.5% of cells) was medium in qualitative assessment (MESF 33,782). Contrastingly, in BCWM.1, CD138 was only expressed on 34.9% of gated cells at a low density (MESF 3,719) and κ light-chain on a minor fraction (26.7%) at a very low density (MESF 633.4). CD38 expression was most prominent in RPCI-WM1, followed by BCWM.1 (91.5% of cells, MESF 9,367.9) and least in MWCL-1 (44.4% of cells, MESF 2,108) (Table 3). In contrast to RPCI-WM1 cells, CD28 was expressed in <20% of BCWM.1 or MWCL-1 cells and whose surface density was very low. As anticipated, CD10 and 11c were not expressed in either of three cell lines.


Immunophenotyping of Waldenströms macroglobulinemia cell lines reveals distinct patterns of surface antigen expression: potential biological and therapeutic implications.

Paulus A, Chitta KS, Wallace PK, Advani PP, Akhtar S, Kuranz-Blake M, Ailawadhi S, Chanan-Khan AA - PLoS ONE (2015)

WM-specific antigen expression compared across RPCI-WM1, BCWM.1 and MWCL-1 cell lines.Fluorescein (FITC), phycoerythrin (PE), phycoerythrin—cyanine 5 (PC5) or allophycocyanin (APC) conjugates of various antibodies were used as presented above. All cell lines were negative for CD10 and 11c. Blue line indicates RPCI-WM1 antigen expression, red line indicates MWCL-1 antigen expression and green line indicates antigen expression in BCWM.1 cell line. Only BCWM.1 an MWCL-1 were CD19+, CD20+ and FMC7+. Expression of CD138 and κ-light-chain was seen only on MWCL-1 and RPCI-WM1 cells. CD28 expression was markedly more observable on RPCI-WM1 tumor cells as compared to MWCL-1 or BCWM.1.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4390194&req=5

pone.0122338.g002: WM-specific antigen expression compared across RPCI-WM1, BCWM.1 and MWCL-1 cell lines.Fluorescein (FITC), phycoerythrin (PE), phycoerythrin—cyanine 5 (PC5) or allophycocyanin (APC) conjugates of various antibodies were used as presented above. All cell lines were negative for CD10 and 11c. Blue line indicates RPCI-WM1 antigen expression, red line indicates MWCL-1 antigen expression and green line indicates antigen expression in BCWM.1 cell line. Only BCWM.1 an MWCL-1 were CD19+, CD20+ and FMC7+. Expression of CD138 and κ-light-chain was seen only on MWCL-1 and RPCI-WM1 cells. CD28 expression was markedly more observable on RPCI-WM1 tumor cells as compared to MWCL-1 or BCWM.1.
Mentions: We then conducted a comprehensive comparative analysis of surface markers in WM cell lines (only 3 noted in the medical literature, 2 developed by the Mayo Clinic group; MWCL-1 and RPCI-WM1,[10, 14] and 1 developed at Dana Farber Cancer Institute; BCWM.1).[13] Using the same set of 19 antigens (Fig 1) a comparative analysis was performed in BCWM.1 and MWCL-1 (Fig 2). Both MWCL-1 and BCWM.1 cells were CD19+low and CD20+medium. The CD20 epitope, FMC7, was also expressed in ~45–63% of BCWM.1 and MWCL-1 cell, respectively albeit at low levels. Notably both RPCI-WM1 (88.7% of cells) and MWCL-1 (98.3% of cells) were CD138+medium. RPCI-WM1 (99.1% of cells) demonstrated low density of κ light-chain (MESF 10,789.6) whereas density of κ light-chain on MWCL-1 (99.5% of cells) was medium in qualitative assessment (MESF 33,782). Contrastingly, in BCWM.1, CD138 was only expressed on 34.9% of gated cells at a low density (MESF 3,719) and κ light-chain on a minor fraction (26.7%) at a very low density (MESF 633.4). CD38 expression was most prominent in RPCI-WM1, followed by BCWM.1 (91.5% of cells, MESF 9,367.9) and least in MWCL-1 (44.4% of cells, MESF 2,108) (Table 3). In contrast to RPCI-WM1 cells, CD28 was expressed in <20% of BCWM.1 or MWCL-1 cells and whose surface density was very low. As anticipated, CD10 and 11c were not expressed in either of three cell lines.

Bottom Line: RPCI-WM1 cells demonstrated decreased expression of CD19, CD20, and CD23 with enhanced expression of CD28, CD38 and CD184, antigens that were differentially expressed on BCWM.1 and MWCL-1 cells.Due to increased expression of CD184/CXCR4 and CD38, RPCI-WM1 represents a valuable model in which to study the effects anti-CXCR4 or anti-CD38 targeted therapies that are actively being developed for treatment of hematologic cancers.Our analysis defines the utility of the most commonly employed WM cell lines as based on their immunophenotype profiles, highlighting unique differences that can be further studied for therapeutic exploit.

View Article: PubMed Central - PubMed

Affiliation: Department of Cancer Biology, Mayo Clinic, Jacksonville, Florida, United States of America.

ABSTRACT
Waldenströms macroglobulinemia (WM) is a subtype of Non-Hodgkin's lymphoma in which the tumor cell population is markedly heterogeneous, consisting of immunoglobulin-M secreting B-lymphocytes, plasmacytoid lymphocytes and plasma cells. Due to rarity of disease and scarcity of reliable preclinical models, many facets of WM molecular and phenotypic architecture remain incompletely understood. Currently, there are 3 human WM cell lines that are routinely used in experimental studies, namely, BCWM.1, MWCL-1 and RPCI-WM1. During establishment of RPCI-WM1, we observed loss of the CD19 and CD20 antigens, which are typically present on WM cells. Intrigued by this observation and in an effort to better define the immunophenotypic makeup of this cell line, we conducted a more comprehensive analysis for the presence or absence of other cell surface antigens that are present on the RPCI-WM1 model, as well as those on the two other WM cell lines, BCWM.1 and MWCL-1. We examined expression of 65 extracellular and 4 intracellular antigens, comprising B-cell, plasma cell, T-cell, NK-cell, myeloid and hematopoietic stem cell surface markers by flow cytometry analysis. RPCI-WM1 cells demonstrated decreased expression of CD19, CD20, and CD23 with enhanced expression of CD28, CD38 and CD184, antigens that were differentially expressed on BCWM.1 and MWCL-1 cells. Due to increased expression of CD184/CXCR4 and CD38, RPCI-WM1 represents a valuable model in which to study the effects anti-CXCR4 or anti-CD38 targeted therapies that are actively being developed for treatment of hematologic cancers. Overall, differences in surface antigen expression across the 3 cell lines may reflect the tumor clone population predominant in the index patients, from whom the cell lines were developed. Our analysis defines the utility of the most commonly employed WM cell lines as based on their immunophenotype profiles, highlighting unique differences that can be further studied for therapeutic exploit.

No MeSH data available.


Related in: MedlinePlus