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Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection.

Rosén A, Bergh AC, Gogok P, Evaldsson C, Myhrinder AL, Hellqvist E, Rasul A, Björkholm M, Jansson M, Mansouri L, Liu A, Teh BT, Rosenquist R, Klein E - Oncoimmunology (2012)

Bottom Line: Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments.These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment.Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine; Division of Cell Biology; Linköping University; Linköping, Sweden.

ABSTRACT
Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1-2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16-1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

No MeSH data available.


Related in: MedlinePlus

Figure 6. Multicolor immunofluorescence for EBNA-2 and LMP-1. From left to right: DAPI staining (Vectashield) for nuclear DNA; EBNA-2 staining with mouse anti-EBNA-2 mAb clone PE2, followed by goat anti-mouse IgG1-Alexa488; LMP-1 staining with anti-LMP-1 mAb clone S-12, followed by anti-mouse IgG-Alexa594; Merged image of EBNA-2 and LMP-1.
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Figure 6: Figure 6. Multicolor immunofluorescence for EBNA-2 and LMP-1. From left to right: DAPI staining (Vectashield) for nuclear DNA; EBNA-2 staining with mouse anti-EBNA-2 mAb clone PE2, followed by goat anti-mouse IgG1-Alexa488; LMP-1 staining with anti-LMP-1 mAb clone S-12, followed by anti-mouse IgG-Alexa594; Merged image of EBNA-2 and LMP-1.

Mentions: The EBV encoded nuclear antigen EBNA-2 and the latent membrane protein LMP-1 were expressed in all cells as seen by immunofluorescence. This characterizes the growth program denoted as type III EBV expression. As in LCLs generated from normal B lymphocyte population, the levels of both proteins were variable and they were not related. Figure 6 shows the simultaneous staining with both antibodies. Similar results were detected by single staining for each antigen (not shown).


Lymphoblastoid cell line with B1 cell characteristics established from a chronic lymphocytic leukemia clone by in vitro EBV infection.

Rosén A, Bergh AC, Gogok P, Evaldsson C, Myhrinder AL, Hellqvist E, Rasul A, Björkholm M, Jansson M, Mansouri L, Liu A, Teh BT, Rosenquist R, Klein E - Oncoimmunology (2012)

Figure 6. Multicolor immunofluorescence for EBNA-2 and LMP-1. From left to right: DAPI staining (Vectashield) for nuclear DNA; EBNA-2 staining with mouse anti-EBNA-2 mAb clone PE2, followed by goat anti-mouse IgG1-Alexa488; LMP-1 staining with anti-LMP-1 mAb clone S-12, followed by anti-mouse IgG-Alexa594; Merged image of EBNA-2 and LMP-1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3376971&req=5

Figure 6: Figure 6. Multicolor immunofluorescence for EBNA-2 and LMP-1. From left to right: DAPI staining (Vectashield) for nuclear DNA; EBNA-2 staining with mouse anti-EBNA-2 mAb clone PE2, followed by goat anti-mouse IgG1-Alexa488; LMP-1 staining with anti-LMP-1 mAb clone S-12, followed by anti-mouse IgG-Alexa594; Merged image of EBNA-2 and LMP-1.
Mentions: The EBV encoded nuclear antigen EBNA-2 and the latent membrane protein LMP-1 were expressed in all cells as seen by immunofluorescence. This characterizes the growth program denoted as type III EBV expression. As in LCLs generated from normal B lymphocyte population, the levels of both proteins were variable and they were not related. Figure 6 shows the simultaneous staining with both antibodies. Similar results were detected by single staining for each antigen (not shown).

Bottom Line: Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments.These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment.Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Experimental Medicine; Division of Cell Biology; Linköping University; Linköping, Sweden.

ABSTRACT
Chronic lymphocytic leukemia (CLL) cells express the receptor for Epstein-Barr virus (EBV) and can be infected in vitro. Infected cells do not express the growth-promoting set of EBV-encoded genes and therefore they do not yield LCLs, in most experiments. With exceptional clones, lines were obtained however. We describe a new line, HG3, established by in vitro EBV-infection from an IGHV1-2 unmutated CLL patient clone. All cells expressed EBNA-2 and LMP-1, the EBV-encoded genes pivotal for transformation. The karyotype, FISH cytogenetics and SNP-array profile of the line and the patient's ex vivo clone showed biallelic 13q14 deletions with genomic loss of DLEU7, miR15a/miR16-1, the two micro-RNAs that are deleted in 50% of CLL cases. Further features of CLL cells were: expression of CD5/CD20/CD27/CD43 and release of IgM natural antibodies reacting with oxLDL-like epitopes on apoptotic cells (cf. stereotyped subset-1). Comparison with two LCLs established from normal B cells showed 32 genes expressed at higher levels (> 2-fold). Among these were LHX2 and LILRA. These genes may play a role in the development of the disease. LHX2 expression was shown in self-renewing multipotent hematopoietic stem cells, and LILRA4 codes for a receptor for bone marrow stromal cell antigen-2 that contributes to B cell development. Twenty-four genes were expressed at lower levels, among these PARD3 that is essential for asymmetric cell division. These genes may contribute to establish precursors of CLL clones by regulation of cellular phenotype in the hematopoietic compartment. Expression of CD5/CD20/CD27/CD43 and spontaneous production of natural antibodies may identify the CLL cell as a self-renewing B1 lymphocyte.

No MeSH data available.


Related in: MedlinePlus