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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 4. Zoom in picture of the homozygous 13q4 deletion. The array revealed that DLEU-7, miR16–1 and miR15a were deleted.
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Figure 4: Figure 4. Zoom in picture of the homozygous 13q4 deletion. The array revealed that DLEU-7, miR16–1 and miR15a were deleted.

Mentions: The karyotypes of the ex vivo cells and the cell line showed biallelic 13q14 deletions, 46,XY,del(13)(q14)x2 (Fig. 3). It was confirmed in fluorescence in situ hybridization (FISH) and single nucleotide polymorphism (SNP) array analysis. The biallelic deletions included genomic loss of DLEU7, miR15a and miR16–1 (Fig. 4). Real-time quantitative polymerase chain reaction (RQ-PCR) confirmed loss of miR15a expression, however, miR16–1 RQ-PCR was positive. In addition, insertions were detected on chromosomes 11 and 14 in the cell line (Fig. 3A and B) but not in the ex vivo CLL cells.


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 4. Zoom in picture of the homozygous 13q4 deletion. The array revealed that DLEU-7, miR16–1 and miR15a were deleted.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Figure 4. Zoom in picture of the homozygous 13q4 deletion. The array revealed that DLEU-7, miR16–1 and miR15a were deleted.
Mentions: The karyotypes of the ex vivo cells and the cell line showed biallelic 13q14 deletions, 46,XY,del(13)(q14)x2 (Fig. 3). It was confirmed in fluorescence in situ hybridization (FISH) and single nucleotide polymorphism (SNP) array analysis. The biallelic deletions included genomic loss of DLEU7, miR15a and miR16–1 (Fig. 4). Real-time quantitative polymerase chain reaction (RQ-PCR) confirmed loss of miR15a expression, however, miR16–1 RQ-PCR was positive. In addition, insertions were detected on chromosomes 11 and 14 in the cell line (Fig. 3A and B) but not in the ex vivo CLL cells.

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