1987 Fiscal Year Final Research Report Summary
Establishment of an antigen specific B cell clone by somatic hybridization
| Project/Area Number |
60570229
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Hyogo College of Medicine |
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Principal Investigator |
HAMANO Teruaki Department of Internal Medicine,Hyogo College of Medicine, 医学部, 講師 (40068515)
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| Co-Investigator(Kenkyū-buntansha) |
IWASAKI Tsuyoshi Department of Internal Medicine,Hyogo College of Medicine, 医学部, 助手 (10151721)
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| Project Period (FY) |
1985 – 1987
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| Keywords | an antigen specific B cell clone / TNP-PFC / TNP-RFC / B細胞表面抗原 |
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| Research Abstract |
Splenic B cells of A/J mice immunized with 2,4,6 trinitrophenyl(TNP)lipopolysaccharide were fused with 2.52M,a mutant of a B cell line,in the presence of polyethylene glycol and dimethyl sulfoxide.TP67.21,a subline of a resulting hybridoma,expresses IA^k,IE^k,IgM,B220, P50,and receptors for C3 fragment of complement,the Fc portion of IgG,and interleuken 2 receptor on the cell membrane;it also possesses receptor molecules for TNP on its surface,derived from TNP reactive B cells of A/J mice primed with TNP LPS used for somatic hybridization,by a rosette forming assay with TNP-SRBC.In contrast, parental 2.52M lacks IA^k and IE^k on the cell membrane and dose not bind to TNP SRBC under the same conditions.Thus,it is likely that TP67.21 is and antigen specific B cell clone directly against TNP.The antigen binding of cells was markedly inhibited by the specific free hapten or anti IgM antibodies.Interestingly,TP67.21 was induced to generate a significant amount of anti TNP antibody when treated with TNP conjugates including T cell independent and dependent antigens,such as TNP-LPS,TNP-BSA,TNP-OVA, and TNP-KLH in the absence of T cell help,as well as polyclonal actibators;this was followed by a marked decrease in the expression of B cell surface markers on the cell membrane.This suggests that the crosslinkage of receptor molecules on TP67.21 by antigen may directly provide a differentiative signal for maturation to a lineage of B cells, and consequently results in the generantion of antigen specific antibodies without T cell involvement.
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