Tohoku
J. Exp.
Med., 1991, 163, 289-294
B-Lineage
Phenotype
Lymphoblastoid Patients
with
of
Cell Lines
from
X-Linked
Agammaglobulinemia SHIGERUTSUCHIYA,HIROMI FUJIE and TASUKEKONNO Department of Pediatrics, the Research Institute for Tuberculosis and Cancer, Tohoku University, Sendai 980
TSUCHIYA, S., FUJIE, H. and KONNO,T. B-Lineage Phenotype of Lymphoblastoid Cell Lines from Patients with X-Linked Agammaglobulinemia. Tohoku J. Exp. Med., 1991, 163 (4), 289-294 Epstein-Barr virus (EBV)-induced precursor B-cell lines were established from bone marrow cells of patients with X-linked agammaglobulinemia (X-LA). Using seven B-lineage specific monoclonal antibodies marker profiles of these cell lines were examined in order to know developmental stage specific and/or X-LA specific changes of B-cell related cellsurface antigens. CD19, CD20, CR2 (CD21), Fc E receptor (CD23) and HLA-DR antigens were consistently found on the X-LA derived precursor B-cell lines as well as mature lymphoblastoid cell lines from healthy adults. These results suggest that immortalisation of B-cells with EBV is always accompanied by expression of pan-B cell markers and B-cell activation antigen (Fc s receptor) even though the cell lines have precursor B-cell phenotypes as defined by immunoglobulin expression. X-linked agammaglobulinemia ; bone marrow ; precursor B cell lines ; B-lineage phenotype
It is extremely difficult to handle normal precursor B cells directly. Usually, HLA-DR positive leukemia/lymphoma cells with or without CD10 antigen are used as being representative of precursor B cells, because of the finding of immunoglobulin gene rearrangements (Korsmeyer et al. 1983). Through analyses of surface antigens of those cells a hypothetical model of normal B-cell development has been proposed as the orderly acquisition of B-cell associated antigens (Anderson et al. 1984). According to this model, HLA-DR, CD19, CD20 and CR2 (CD21) are expressed in this order. Epstein-Barr virus (EBV) induced B-lymphoblastoid cell lines (B-LCL) are another source of human precursor B-cells. X-linked agammaglobulinemia (XLA) is characterized by the absence of circulating B cells and the presence of pre-B cells in bone marrow with a normal frequency. Since there are no mature B-cells in the bone marrow of X-LA patients, X-LA derived precursor B-cells are easily immortalized from bone marrow cells by infection with EBV (Fu et al. Received
December
4, 1990;
revision
accepted 289
for publication
March
22, 1991.
290
S. Tsuchiya
et al.
1980; Tsuchiya et al. 1983 ; Levitt et al. 1984 ; Kubagawa et al. 1988 ; Lau et al. 1989). Development of B-cells is stopped at the same immunological stage as in the fetal liver, and we have shown that the fetal liver is also the source of normal precursor B-cells without X-LA background (Katamine et al. 1984). Several EBV-induced immature B-cell lines have been established in our laboratory from the bone marrow of X-LA patients and the pattern of immunoglobulin expression of these cell lines has already been reported (Tsuchiya et al. 1983). However, it is still unknown whether X-LA derived precursor B-cell lines express B-cell associated surface antigens according to the rule observed on leukemia and lymphoma cells. In this communication we examined the expression of known B-cell associated cell surface antigens on X-LA derived precursor B-cell lines in order to know developmental stage specific and X-LA specific changes of cell surface antigens. MATERIALS AND METHODS Patients Immunological data of three boys with agammaglobulinemia (K.Ko., M.O. and OS.) has been described previously (Tsuchiya et al. 1983). The diagnosis of X-linked agammaglobulinemia (X-LA) was based on the following criteria : 1) male sex, 2) onset in infancy or early childhood, 3) serum IgG
J. Exp.
Med., 1991, 163, 289-294
B-Lineage
Phenotype
Lymphoblastoid Patients
with
of
Cell Lines
from
X-Linked
Agammaglobulinemia SHIGERUTSUCHIYA,HIROMI FUJIE and TASUKEKONNO Department of Pediatrics, the Research Institute for Tuberculosis and Cancer, Tohoku University, Sendai 980
TSUCHIYA, S., FUJIE, H. and KONNO,T. B-Lineage Phenotype of Lymphoblastoid Cell Lines from Patients with X-Linked Agammaglobulinemia. Tohoku J. Exp. Med., 1991, 163 (4), 289-294 Epstein-Barr virus (EBV)-induced precursor B-cell lines were established from bone marrow cells of patients with X-linked agammaglobulinemia (X-LA). Using seven B-lineage specific monoclonal antibodies marker profiles of these cell lines were examined in order to know developmental stage specific and/or X-LA specific changes of B-cell related cellsurface antigens. CD19, CD20, CR2 (CD21), Fc E receptor (CD23) and HLA-DR antigens were consistently found on the X-LA derived precursor B-cell lines as well as mature lymphoblastoid cell lines from healthy adults. These results suggest that immortalisation of B-cells with EBV is always accompanied by expression of pan-B cell markers and B-cell activation antigen (Fc s receptor) even though the cell lines have precursor B-cell phenotypes as defined by immunoglobulin expression. X-linked agammaglobulinemia ; bone marrow ; precursor B cell lines ; B-lineage phenotype
It is extremely difficult to handle normal precursor B cells directly. Usually, HLA-DR positive leukemia/lymphoma cells with or without CD10 antigen are used as being representative of precursor B cells, because of the finding of immunoglobulin gene rearrangements (Korsmeyer et al. 1983). Through analyses of surface antigens of those cells a hypothetical model of normal B-cell development has been proposed as the orderly acquisition of B-cell associated antigens (Anderson et al. 1984). According to this model, HLA-DR, CD19, CD20 and CR2 (CD21) are expressed in this order. Epstein-Barr virus (EBV) induced B-lymphoblastoid cell lines (B-LCL) are another source of human precursor B-cells. X-linked agammaglobulinemia (XLA) is characterized by the absence of circulating B cells and the presence of pre-B cells in bone marrow with a normal frequency. Since there are no mature B-cells in the bone marrow of X-LA patients, X-LA derived precursor B-cells are easily immortalized from bone marrow cells by infection with EBV (Fu et al. Received
December
4, 1990;
revision
accepted 289
for publication
March
22, 1991.
290
S. Tsuchiya
et al.
1980; Tsuchiya et al. 1983 ; Levitt et al. 1984 ; Kubagawa et al. 1988 ; Lau et al. 1989). Development of B-cells is stopped at the same immunological stage as in the fetal liver, and we have shown that the fetal liver is also the source of normal precursor B-cells without X-LA background (Katamine et al. 1984). Several EBV-induced immature B-cell lines have been established in our laboratory from the bone marrow of X-LA patients and the pattern of immunoglobulin expression of these cell lines has already been reported (Tsuchiya et al. 1983). However, it is still unknown whether X-LA derived precursor B-cell lines express B-cell associated surface antigens according to the rule observed on leukemia and lymphoma cells. In this communication we examined the expression of known B-cell associated cell surface antigens on X-LA derived precursor B-cell lines in order to know developmental stage specific and X-LA specific changes of cell surface antigens. MATERIALS AND METHODS Patients Immunological data of three boys with agammaglobulinemia (K.Ko., M.O. and OS.) has been described previously (Tsuchiya et al. 1983). The diagnosis of X-linked agammaglobulinemia (X-LA) was based on the following criteria : 1) male sex, 2) onset in infancy or early childhood, 3) serum IgG
