Induction of Ly-49 on an interspecies hybridoma between differentiation stage specific murine T cells and a rat T lymphoma.
Ly-49 can be expressed in interspecies hybridoma cells resulting from the fusion of Ly-49 negative lymphoid cells from normal mice and Ly-49 negative T cells from the rat thymoma W/Fu(C58NT)D (C58). Several hundred hybridoma clones of this type were studied. Hybridoma clones derived from the fusion between activated C57BL/6 mouse spleen cells and C58 expressed Ly-49 on their surface although Ly-49 was undetectable on both parental cells. No Ly-49 antigen was expressed on hybrids when BW5147, a T lymphoma derived from AKR mouse, replaced C58 as parental cell. Thymocytes from C57BL/6 mice also produced Ly-49 positive interspecies hybridomas with C58. Subsequent manipulation of these thymocytes with a mixture of anti-CD4 and anti-CD8 antibodies plus complement to obtain double negative cells (CD4-/CD8-) prior to the fusion abolished the expression of Ly-49 on hybridomas. On the other hand, if the thymocytes were treated with either anti-CD4 or anti-CD8 antibody plus complement, the resultant thymocytes which were a mixture of cells with a single positive (CD4+/CD8- or CD4-/CD8+) and a double negative phenotype (CD4-/CD8-) retained the ability to produce Ly-49 positive hybridomas. Interspecies hybridomas between either CD4+/CD8- or CD4-/CD8+ functional T cell clones and C58 expressed Ly-49 on their surface, though these parental T cells lacked Ly-49 on their surfaces. Northern hybridization analysis using Ly-49 cDNA revealed that C58 cells do not express Ly-49 mRNA, indicating that a trans-acting factor(s) from C58 plays an important role in the induction of cell surface Ly-49 expression on interspecies hybridomas. Our data indicate the complex nature of the regulatory mechanisms for expression of the Ly-49 molecule.
Nagasawa
R
R
Department of Internal Medicine, Saitama Medical Center, Saitama Medical School, Japan.
Maruyama
N
N
Kubo
S
S
Yumura
W
W
Mitarai
T
T
Isoda
K
K
Kanagawa
O
O
D000818:Animals; D000950:Antigens, Ly / Q000235:genetics / Q000276:immunology*; D002454:Cell Differentiation; D015870:Gene Expression; D006825:Hybridomas / Q000276:immunology*; D037181:Lectins, C-Type; D008213:Lymphocyte Activation; D016399:Lymphoma, T-Cell / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D055630:Receptors, NK Cell Lectin-Like; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology*
D000950:Antigens, Ly; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D055630:Receptors, NK Cell Lectin-Like