In vivo CTL induction with point-substituted ovalbumin peptides: immunogenicity correlates with peptide-induced MHC class I stability

doi:10.1016/0264-410X(95)93320-9

Vaccine

Volume 13, Issue 3, 1995, Pages 313-320

https://doi.org/10.1016/0264-410X(95)93320-9 Get rights and content

Abstract

Class I molecules are conformationally sensitive to peptide binding, prolonging the complex's half-life on the surface of the cell. By making a series of H2K^b anchor motif amino acid point substitutions in the ovalbumin 257–264 octamer, we were able to analyse subtle changes in peptide binding, K^b stabilization and in vivo immunogenicity. The cell line RMA-S was used to determine peptide-dependent K^b stabilization under equilibrium and non-equilibrium binding conditions. Sixteen conservative and non-conservative amino acid substitutions were made at positions 3, 5 or 8 of the peptide. At 37°C, K^b stabilization was differentially affected by these substitutions, with several substitutions severely affecting K^b surface expression. When the substituted peptides were used as immunogens to prime cytotoxic T lymphocytes (CTL) in vivo, each peptide's ability to stabilize K^b directly correlated with the intensity of specific CTL activation. We conclude that peptide class I stabilization is an important influencing factor in determining cell surface steady-state expression of these peptides and thus the breadth of CTL recruitment. These concepts may relate the phenomenon of immunodominance to cell surface-presented peptide steady-state levels and may also aid in peptide vaccine design.

References (33)

E.A. Stura et al.
Crystallization of murine major histocompatibility complex class I H-2Kb with single peptides
J. Mol. Biol.
(1992)
G.B. Lipford et al.
The immunodominant peptide from listerolysin in Quil A liposomes vaccinates CD8 + CTL and confers protection to infection
Immunol. Lett.
(1994)
G.B. Lipford et al.
Vaccination with immunodominant peptides encapsulated in Quil A-containing liposomes induces peptide-specific primary CD8 + cytotoxic T cells
Vaccine
(1994)
A. Townsend et al.
Antigen recognition by class I-restricted T lymphocytes
Annu. Rev. Immunol.
(1989)
P.J. Bjorkman et al.
Structure, function, and diversity of class I major histocompatility complex molecules
Annu. Rev. Biochem.
(1990)
T.P. Garrett et al.
Specificity pockets for the side chains of peptide antigens in HLA-Aw68
Nature
(1989)
T.S. Jardetzky et al.
Identification of self peptides bound to purified HLA-B27
Nature
(1991)
D.H. Fremont et al.
Crystal structures of two viral peptides in complex with murine MHC class I H-2Kb
Science
(1992)
M. Matsumura et al.
Emerging principles for the recognition of peptide antigens by MHC class I molecules
Science
(1992)
K. Falk et al.
Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules
Nature
(1991)

E.G. Pamer et al.

Precise prediction of a dominant class I MHC-restricted epitope of Listeria monocytogenes

Nature

(1991)

G.B. Lipford et al.

Primary in vivo responses to ovalbumin. Probing the predictive value of the Kb binding motif

J. Immunol.

(1993)

G.M. Van Bleek et al.

The structure of the antigen-binding groove of major histocompatibility complex class I molecules determines specific selection of self-peptides

M.R. Lie et al.

The specific binding of peptide ligand to Ld class I major histocompatibility complex molecules determines their antigenic structure

J. Exp. Med.

(1991)

W.R. Lie et al.

Peptide ligand-induced conformation and surface expression of the Ld class I MHC molecule

Nature

(1990)

T. Elliott et al.

Peptide-induced conformational change of the class I heavy chain

Nature

(1991)

Cited by (48)

A deterministic model for the processing and presentation of bacteria-derived antigenic peptides
2008, Journal of Theoretical Biology
The amount and the dynamics of antigen supply to the cellular antigen processing and presentation machinery differ largely among diverse microbial antigens and various types of antigen presenting cells. The precise influence, however, of antigen supply on the antigen presentation pattern of cells is not known. Here, we provide a basic deterministic mathematical model of antigen processing and presentation of microbial antigens. The model predicts that different types of antigen presenting cells e.g. cells presenting or cross-presenting exogenous antigens, cells infected with replicating microbes, or cells in which microbial antigen synthesis is blocked after a certain period of time have inherently different antigen presentation patterns which are defined by the kinetics of antigen supply. The reevaluation of existing experimental data [Sijts, A.J., Pamer, E.G., 1997. Enhanced intracellular dissociation of major histocompatibility complex class I-associated peptides: a mechanism for optimizing the spectrum of cell surface-presented cytotoxic T lymphocyte epitopes. J. Exp. Med. 185, 1403–1411] describing the processing and presentation of two antigenic peptides derived from the p60 proteins of the facultatively intracellular bacterium Listeria monocytogenes shows that p60 proteins accumulating intracellularly during bacterial infection of cells play no measurable role as substrate for the cytosolic antigen presentation pathway.
Cytotoxic T cells generated against heteroclitic peptides kill primary tumor cells independent of the binding affinity of the native tumor antigen peptide
2006, Blood
Citation Excerpt :
To date it remains controversial as to how reliable the predicted binding corresponds to experimental binding and to what extent the binding affinity translates into immunogenicity and subsequent generation of effective CTLs. Several reports analyzing non-Ig–derived peptides have demonstrated poor correlation between predicted and actual binding as determined by peptide-binding assay or by complex stabilization.45-50 Our own results point to the increasing efficiency of matrix-driven epitope deduction analyzing Ig-derived peptides because Rammensee scores, Parker scores, and binding in the T2 binding assay correlated highly.
Heteroclitic peptide modifications increase immunogenicity, allowing generation of cytotoxic T lymphocytes (CTLs) against weakly immunogenic tumor-associated antigens (TAAs). A critical issue is whether T cells generated against heteroclitic peptides retain the ability to recognize and kill tumor cells expressing the original weak TAAs, and whether there is a lower threshold of binding affinity of the native peptides, below which such CTLs can still kill primary tumor cells. To examine this we used a model examining the ability of native and heteroclitic immunoglobulin (Ig)–derived peptides to generate CTLs that can kill chronic lymphocytic leukemia (CLL) cells. We demonstrate that CTLs generated against heteroclitic peptides have enhanced killing of CD40-activated B cells pulsed with either heteroclitic (P < .001) or native peptide (P = .04) and primary CLL cells (P = .01). The novel finding reported here is that the rate-limiting factor appears to be the ability to generate CTLs and that once generated, CTL lysis of primary tumor cells is independent of the binding affinity of the native peptide. These findings have implications for vaccination strategies in malignancies and are currently being further examined in vivo in murine models.
Functional and structural characteristics of NY-ESO-1-related HLA A2-restricted epitopes and the design of a novel immunogenic analogue
2004, Journal of Biological Chemistry
Citation Excerpt :
The C9Abu analogue was consistently recognized more efficiently by the T cell lines, and as a general rule the following reactivity pattern was observed: C9Abu > C9A, C9V > wild type > C9S > C9L. This did not simply correlate directly with binding or stability of the complexes, as may be expected (6, 50-57), and is consistent with several other studies that show that the immunogenicity of some T cell determinants is influenced by additional factors (9, 58-62). C9S bound to HLA A2 slightly less efficiently than the wild type peptide yet demonstrated drastically worse stabilization of the complexes.
NY-ESO-1, a commonly expressed tumor antigen of the cancer-testis family, is expressed by a wide range of tumors but not found in normal adult somatic tissue, making it an ideal cancer vaccine candidate. Peptides derived from NY-ESO-1 have shown preclinical and clinical trial promise; however, biochemical features of these peptides have complicated their formulation and led to heterogeneous immune responses. We have taken a rational approach to engineer an HLA A2-restricted NY-ESO-1-derived T cell epitope with improved formulation and immunogenicity to the wild type peptide. To accomplish this, we have solved the x-ray crystallographic structures of HLA A2 complexed to NY-ESO (157-165) and two analogues of this peptide in which the C-terminal cysteine residue has been substituted to alanine or serine. Substitution of cysteine by serine maintained peptide conformation yet reduced complex stability, resulting in poor cytotoxic T lymphocyte recognition. Conversely, substitution with alanine maintained complex stability and cytotoxic T lymphocyte recognition. Based on the structures of the three HLA A2 complexes, we incorporated 2-aminoisobutyric acid, an isostereomer of cysteine, into the epitope. This analogue is impervious to oxidative damage, cysteinylation, and dimerization of the peptide epitope upon formulation that is characteristic of the wild type peptide. Therefore, this approach has yielded a potential therapeutic molecule that satiates the hydrophobic F pocket of HLA A2 and exhibited superior immunogenicity relative to the wild type peptide.
Induction of a cytotoxic T-cell response to HIV-1 proteins with short synthetic peptides and human compatible adjuvants
2001, Vaccine
The goal of this study was the induction of a strong CTL response against multiple CTL epitopes present in HIV proteins using short synthetic peptides. Four HLA-A2.1 restricted peptides (RT 476–484, p17 77–85, gp41 814–823, RT 956–964) that showed stable binding to the HLA-A2.1 molecule in an in vitro binding assay were able to elicit a strong specific immune response in HLA-A2.1 transgenic mice when injected with IFA or Montanide®. The use of biodegradable microspheres (MS) as adjuvant was also successfully tested for all peptides. When the peptides were injected as a mixture the response was weaker as compared to individual injections of the peptides indicating the occurrence of immunodominance (ID). We are currently investigating whether ID can be overcome by a combined injection of peptide loaded MS with different release patterns. Taken together, it seems feasible to induce a specific CTL response in HLA-A2.1 transgenic mice against several HIV proteins using short synthetic peptides and human compatible adjuvants.
Evaluation of T-cell responses to peptides and lipopeptides with MHC class I binding motifs derived from the amino acid sequence of the 19-kDa lipoprotein of mycobacterium tuberculosis
2000, Molecular Immunology
Cytotoxic T-lymphocyte (CTL) epitopes on the 19-kDa lipoprotein from Mycobacterium tuberculosis were identified by the use of lipopeptides and their cytokine profile studied. Selection of candidate CTL epitopes was based on synthetic peptides derived from the amino acid sequence of the 19-kDa lipoprotein showing major histocompatibility complex class I (MHC-I) binding motifs (H-2D^b and H-2L^d). Their ability to up-regulate and stabilize MHC-I molecules on the mouse lymphoma cell line RMA-S was studied. Similar studies were performed with peptides, in which the anchor amino acid of the H-2D^b MHC-I motif was replaced by alanine. Three out of five peptides with H-2D^b or H-2L^d binding motifs and their corresponding lipopeptides as well, up-regulated and stabilized the H-2D^b molecules on RMA-S cells. Replacement of the anchor amino acid residues of the H-2D^b MHC-I motif by alanine revealed that the anchor amino acid asparagine at position 5, contributed more to binding of peptide to H-2D^b molecules than leucine at position 11. The closely related lipopeptides LP19c and LP19d, in combination with incomplete Freund's adjuvant (IFA), induced CTL responses in C57BL/6 (H-2^b) mice. These CTLs could recognize the naturally processed antigen, i.e. the 19-kDa antigen protein produced and processed by the EX-19 cell line. The capacity of the various lipopeptides to induce CTL correlated well with the ability of the (lipo)peptide to up-regulate and to stabilize H-2D^b molecules. Lipopeptide LP19c primed spleen cells showed a T helper type one profile after in vitro stimulation with P19c and P19d 19 kDa peptides. The approach to characterize presumptive 19-kDa CTL epitopes might lead to selection of promising CTL epitopes, which can be applied in the development of subunit tuberculosis vaccines.
K-ras peptide mimotope induces a humoral immune response against g12v k-ras antigen in balb/c mice
2020, Asia-Pacific Journal of Molecular Biology and Biotechnology

View all citing articles on Scopus

View full text

PaperIn vivo CTL induction with point-substituted ovalbumin peptides: immunogenicity correlates with peptide-induced MHC class I stability

Abstract

J. Mol. Biol.

Immunol. Lett.

Vaccine

Antigen recognition by class I-restricted T lymphocytes

Annu. Rev. Immunol.

Structure, function, and diversity of class I major histocompatility complex molecules

Annu. Rev. Biochem.

Specificity pockets for the side chains of peptide antigens in HLA-Aw68

Nature

Identification of self peptides bound to purified HLA-B27

Nature

Crystal structures of two viral peptides in complex with murine MHC class I H-2Kb

Science

Emerging principles for the recognition of peptide antigens by MHC class I molecules

Science

Allele-specific motifs revealed by sequencing of self-peptides eluted from MHC molecules

Nature

Precise prediction of a dominant class I MHC-restricted epitope of Listeria monocytogenes

Nature

Primary in vivo responses to ovalbumin. Probing the predictive value of the Kb binding motif

J. Immunol.

The structure of the antigen-binding groove of major histocompatibility complex class I molecules determines specific selection of self-peptides

The specific binding of peptide ligand to Ld class I major histocompatibility complex molecules determines their antigenic structure

J. Exp. Med.

Peptide ligand-induced conformation and surface expression of the Ld class I MHC molecule

Nature

Peptide-induced conformational change of the class I heavy chain

Nature

Paper
In vivo CTL induction with point-substituted ovalbumin peptides: immunogenicity correlates with peptide-induced MHC class I stability