Major histocompatibility complex independent T cell receptor-antigen interaction: functional analysis using fluorescein derivatives.
We have isolated T cell receptor (TCR) cDNAs from fluorescein (FL)-specific human T cell clones (alpha FL beta FL), and transferred them to TCR beta- Jurkat cells in order to study direct FL-binding to the TCR. Using either FL-conjugated polymers (FL-polymer) or FL-substituted Sepharose beads, we are able to demonstrate the direct binding of antigen to the T cell surface, and the functional activation of the T cell transfectants. We present evidence against the involvement of major histocompatibility complex (MHC) molecules or antigen presentation in the interaction of FL with the alpha FL beta FL transfectants. Additionally, we have examined the effect of ring substitutions on the FL molecule as well as specific alterations of substituents attached to the 5' position, and we have found that all of them interfere with the functional recognition of the alpha FL beta FL TCR. These experiments demonstrate that TCRs like antibodies have intrinsic affinities for antigen, even without the involvement of MHC molecules.
Diamond
D J
DJ
Division of Immunology, Beckman Research Institute of City of Hope, Duarte, California 91010.
Szalay
P
P
Symer
D
D
Hao
P
P
Shin
H S
HS
Dintzis
R Z
RZ
Dintzis
H M
HM
Reinherz
E L
EL
Siliciano
R F
RF
D000595:Amino Acid Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005452:Fluoresceins; D014644:Genetic Variation; D005822:Genetic Vectors; D006801:Humans; D046911:Macromolecular Substances; D008285:Major Histocompatibility Complex; D008969:Molecular Sequence Data; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000502:physiology; D015183:Restriction Mapping; D012685:Sepharose; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
D005452:Fluoresceins; D046911:Macromolecular Substances; D011948:Receptors, Antigen, T-Cell; D012685:Sepharose