Antigen presentation by Ia+ B cell hybridomas to H-2-restricted T cell hybridomas.

Kappler, John W.; White, Janicé; Wegmann, Dale; Mustain, Eric; Marrack, Philippa

doi:10.1073/pnas.79.11.3604

Cited by 225 publications

(105 citation statements)

References 19 publications

Supporting

Mentioning

102

Contrasting

Unclassified

Order By: Relevance

“…p3K and its variants were synthesized in the Molecular Resource Center of Nation Jewish Medical and Research Center. The IA b -expressing B cell hybridoma, LB-15.13 (35), was used as an antigen-presenting cell for these peptides. Antigen-induced IL-2 release from the T cell hybridomas was assayed as described (36,37).…”

Section: Methodsmentioning

confidence: 99%

Alternate interactions define the binding of peptides to the MHC molecule IA^b

Liu

Dai

Crawford

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

T he crystal structures of a variety of MHCI (1-9) and MHCII (10-23) molecules have revealed their features that lead to stable peptide binding. Conserved amino acids of MHCI interact with the N terminus and C terminus of the peptide in virtually all isotypes and alleles. Between these termini, the peptide can vary in length and in the path it takes through the binding groove, often bulging out of the groove in the middle. In MHCII, the N and C termini of the peptide generally extend beyond the binding groove and do not interact with the MHCII molecule. Rather, H-bonding interactions between the peptide backbone and conserved amino acids of the MHCII ␣-helices force the peptide to take a similar, extended path through the groove of all MHCII isotypes and alleles.The unique interactions that determine the allelic specificity of peptide binding seem to be controlled by the character of pockets within the interior of the binding groove that preferentially accept particular amino acid side chains. Because the MHC amino acids that line these pockets are among the most genetically variable, their depth, shape, and chemistry vary considerably among MHC alleles and isotypes, and they appear to be the most important factor in the specificity of peptide binding.In the current study we have solved the crystal structure of the mouse MHCII molecule IA b with a bound immunogenic peptide variant of a dominant peptide derived from the MHCII E␣ protein that is found in IA b in some strains of mice (24). Despite the high stability and immunogenicity of this complex, the four usual p1, p4, p6, and p9 peptide side chain binding pockets of IA b are largely unfilled, because alanines occur in the peptide at these positions. Rather, the stability of this peptide͞MHCII complex can be attributed to the combination of the conserved interactions with the peptide backbone as well as extensive unique interactions of the IA b molecule with other peptide amino acids particularly at the N-terminal end and the p7 position of the peptide. These results suggest that IA b can achieve stable peptide binding in more than one way and may account for the previous difficulty in defining the IA b peptidebinding motif. Materials and MethodsPreparation of Soluble IA b . The production of soluble IA b containing a peptide attached to the ␤-chain N terminus via a flexible peptide linker has been described (25). In the original constructions, peptides included both the E␣-derived peptide, ASFEAQGALANIAVDKA (pE␣), which occupies about 10% of natural IA b , and an immunogenic variant, ASFEAQKAK-ANKAVDKA (p3K), in which three positions in the peptide (underlined) had lysines substituted for the E␣ peptide amino acids. For crystallization, the construct was altered to use a shortened form of p3K, FEAKGAKANKAVD, and to shorten the IA b ␣ and ␤ chains to the predicted ends of the ␣2 and ␤2 domains. The construct was cloned into a previously described dual promoter baculovirus transfer vector (26, 27) and introduced into BacVector 3000 version of AcNPV baculovirus ...

show abstract

Section: Methodsmentioning

confidence: 99%

Alternate interactions define the binding of peptides to the MHC molecule IA^b

Liu

Dai

Crawford

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…B10.A and B10.D2 mice were purchased from The Jackson Laboratory (Bar Harbor, ME). LK35.2 cell, a B cell hybridoma that expresses I-A d and I-E k was provided by Dr. P. Marrack (Howard Hughes Medical Institute, Denver, CO) and LB27.4, a B cell hybridoma expressing D b and K b molecules (26), was purchased from American Type Culture Collection (Manassas, VA) as was the EL-4 T cell lymphoma.…”

Section: Mice and Cell Linesmentioning

confidence: 99%

Study of the Mechanism of TCR Antagonism Using Dual-TCR-Expressing T Cells

Yang

Grey

2003

The Journal of Immunology

View full text Add to dashboard Cite

The mechanism of action of TCR antagonists is incompletely understood. T cells expressing two distinct TCRs have been used to test competition for TCR occupancy as a potential mechanism. Previous studies with CD4 T cells showed that an antagonist for one TCR inhibited the response to the other TCR (cross-antagonism), whereas studies with CD8 cells failed to demonstrate cross-antagonism. To determine whether CD4 and CD8 cells were intrinsically different or whether the differences were the result of the use of different effector assays, we studied both CD4 and CD8 dual-TCR-expressing T cells. In the CD4 system, consistent with previous reports, cross-antagonism of proliferation was observed. In the CD8 system, cross-antagonism was observed using proliferation as readout but not when target cell cytolysis was used. These results suggest that different mechanisms may be involved in the inhibition of proliferation and inhibition of cytotoxic effector function, the latter only involving competition for TCR occupancy. Inhibition of proliferation appears to be more complex and other mechanisms such as sequestration of signaling molecules or negative signaling may be involved. The fact that 10- to 20-fold more antagonist was needed to achieve cross-antagonism compared with inhibition of the cognate TCR is consistent with the hypothesis that competition for TCR occupancy is also a major, albeit not sole, mechanism of antagonism of the proliferative responses of CD4 and CD8 cells.

show abstract

“…M g -mercaptoethanol (Sigma). The mouse H-2 d/k B cell lymphoma LK35.2 is described in [30]. LK35.2 transfectants expressing the HEL-specific BCR D1.3 and HyHEL-10 [3] were a kind gift from F. Batista.…”

Section: Cell Culture and Facs Analysismentioning

confidence: 99%

Lowering the affinity between antigen and the B cell receptor can enhance antigen presentation

Brooks

Knight

2004

Eur J Immunol

View full text Add to dashboard Cite

The B cell receptor (BCR) enables antigen-specific B cells to bind, internalize and target antigens for processing into small peptide fragments. These epitopes are then expressed on the plasma membrane in association with MHC class II molecules for recognition by CD4 + T cells. The affinity of the interaction between the BCR and antigen plays an important part in determining T cell epitope generation. In this report we provide evidence that the efficiency of antigen presentation by specific B cells does not need to be directly proportional to antigen/BCR affinity. We show that increased presentation can result from lowering the affinity of the antigen/BCR interaction. This finding suggests a novel mechanism by which B cells can recruit T cell help and obtain survival signals. Activation of these cells may have consequences for the generation of the B cell repertoire.

show abstract

Antigen presentation by Ia+ B cell hybridomas to H-2-restricted T cell hybridomas.

Cited by 225 publications

References 19 publications

Alternate interactions define the binding of peptides to the MHC molecule IA^b

Alternate interactions define the binding of peptides to the MHC molecule IA^b

Study of the Mechanism of TCR Antagonism Using Dual-TCR-Expressing T Cells

Lowering the affinity between antigen and the B cell receptor can enhance antigen presentation

Contact Info

Product

Resources

About

Antigen presentation by Ia+ B cell hybridomas to H-2-restricted T cell hybridomas.

Cited by 225 publications

References 19 publications

Alternate interactions define the binding of peptides to the MHC molecule IAb

Alternate interactions define the binding of peptides to the MHC molecule IAb

Study of the Mechanism of TCR Antagonism Using Dual-TCR-Expressing T Cells

Lowering the affinity between antigen and the B cell receptor can enhance antigen presentation

Contact Info

Product

Resources

About

Alternate interactions define the binding of peptides to the MHC molecule IA^b

Alternate interactions define the binding of peptides to the MHC molecule IA^b