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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 5. Gene expression difference observed by RT-PCR. Comparison between gene expression of LILRA4, FCRLA and CYP1B1 using RT-PCR in the CLL-HG cell line and 2 LCL lines IARC-171 and LCL-3M derived from normal B cells.
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Figure 5: Figure 5. Gene expression difference observed by RT-PCR. Comparison between gene expression of LILRA4, FCRLA and CYP1B1 using RT-PCR in the CLL-HG cell line and 2 LCL lines IARC-171 and LCL-3M derived from normal B cells.

Mentions: Genes expressed with a 2-fold (or over) difference in the HG3 cells included LHX, and LILRA4. Among the relatively low expressed genes were PARD3, FCRLA and TCL1. The LHX2 (LIM homeobox 2) gene was expressed 20-fold higher in HG3 compared with LCL-3M (Table 1). In three of four additional CLL-LCLs (232B4, I83-E95, CI, MEC1), high expression of LHX2 was detected (Table 1). The LILRA4 [leukocyte immunoglobulin-like receptor, subfamily A (with TM domain), member 4] gene also showed higher expression (Fig. 5). Reduced expression was found for FCRLA (cytochrome p450, family 1, subfamily B, polypeptide 1) (Fig. 5andTable 1), CYP1B1 (Fig. 5), the polarity complex gene PARD3 [par-3 partitioning defective 3 homolog (C. elegans)] (Table 1) and TCL1A.


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 5. Gene expression difference observed by RT-PCR. Comparison between gene expression of LILRA4, FCRLA and CYP1B1 using RT-PCR in the CLL-HG cell line and 2 LCL lines IARC-171 and LCL-3M derived from normal B cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 5: Figure 5. Gene expression difference observed by RT-PCR. Comparison between gene expression of LILRA4, FCRLA and CYP1B1 using RT-PCR in the CLL-HG cell line and 2 LCL lines IARC-171 and LCL-3M derived from normal B cells.
Mentions: Genes expressed with a 2-fold (or over) difference in the HG3 cells included LHX, and LILRA4. Among the relatively low expressed genes were PARD3, FCRLA and TCL1. The LHX2 (LIM homeobox 2) gene was expressed 20-fold higher in HG3 compared with LCL-3M (Table 1). In three of four additional CLL-LCLs (232B4, I83-E95, CI, MEC1), high expression of LHX2 was detected (Table 1). The LILRA4 [leukocyte immunoglobulin-like receptor, subfamily A (with TM domain), member 4] gene also showed higher expression (Fig. 5). Reduced expression was found for FCRLA (cytochrome p450, family 1, subfamily B, polypeptide 1) (Fig. 5andTable 1), CYP1B1 (Fig. 5), the polarity complex gene PARD3 [par-3 partitioning defective 3 homolog (C. elegans)] (Table 1) and TCL1A.

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