Differential binding properties of B7-H1 and B7-DC to programmed death-1

doi:10.1016/S0006-291X(03)01257-9

Biochemical and Biophysical Research Communications

Volume 307, Issue 3, 1 August 2003, Pages 672-677

https://doi.org/10.1016/S0006-291X(03)01257-9 Get rights and content

Abstract

Programmed death-1 (PD-1) is a negative regulatory receptor expressed on activated T and B cells. Two ligands for PD-1, B7-H1 (PD-L1) and B7-DC (PD-L2), have been identified, but their binding properties have not been characterized yet. In this study, we generated soluble Ig fusion proteins of these molecules and examined the kinetics and relative affinities of the interactions between B7-H1 or B7-DC and PD-1 by flow cytometry and surface plasmon resonance. The interaction of B7-DC/PD-1 exhibited a 2–6-fold higher affinity and had different association/dissociation kinetics compared with the interaction of B7-H1/PD-1. Our results suggest that the differential binding properties of B7-H1 and B7-DC may be responsible for differential contributions of these two PD-1 ligands to immune responses.

Section snippets

Materials and methods

Constructs. Human PD-1 cDNA was generated by RT-PCR from peripheral blood mononuclear cells using primers shown in Table 1 and subsequently subcloned into a pMKITneo vector, kindly provided by K. Maruyama (Tokyo Medical and Dental University, Tokyo). Constructs for Ig fusion proteins consisting of the extracellular portion of human PD-1, B7-H1, or B7-DC and the hinge-C_H2-C_H3 domains of human IgG1 heavy chain were prepared as described previously [28]. The extracellular portions of each molecule

Characterization of B7-H1Ig, B7-DCIg, and PD-1Ig fusion proteins

To characterize the interactions of B7-H1 and B7-DC with PD-1, we generated soluble fusion proteins consisting of the extracellular region of these molecules and the Fc portion of human IgG1. As shown in Fig. 1, SDS–PAGE analysis showed a single band of 150–170 kDa for B7-H1Ig, B7-DCIg, and PD-1Ig under non-reducing conditions, and each band migrated at approximately half size under reducing conditions. These results indicated homo-dimerization of these Ig fusion proteins by disulfide-linkage,

Discussion

Here, we demonstrated that the relative affinity of PD-1 to B7-DC was 2–6-fold higher than that to B7-H1 as estimated by flow cytometric and SPR analyses. The differences in K_d values obtained by flow cytometry and SPR may result from structural differences between the cell surface molecules on transfectants and the immobilized dimeric forms of Ig fusion proteins on BIAcore sensor. However, a higher affinity of B7-DC/PD-1 interaction than B7-H1/PD-1 interaction was consistently observed in both

Acknowledgements

This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology, and from the Ministry of Health, Labor, and Welfare of Japan.

References (40)

P.S. Linsley et al.
Human B7-1 (CD80) and B7-2 (CD86) bind with similar avidities but distinct kinetics to CD28 and CTLA-4 receptors
Immunity
(1994)
A.V. Collins et al.
The interaction properties of costimulatory molecules revisited
Immunity
(2002)
S. Ikemizu et al.
Structure and dimerization of a soluble form of B7-1
Immunity
(2000)
H. Nishimura et al.
PD-1: an inhibitory immunoreceptor involved in peripheral tolerance
Trends Immunol
(2001)
H. Nishimura et al.
Development of lupus-like autoimmune diseases by disruption of the PD-1 gene encoding an ITIM motif-carrying immunoreceptor
Immunity
(1999)
H. Akiba et al.
Identification of rat OX40 ligand by molecular cloning
Biochem. Biophys. Res. Commun
(1998)
H. Kozono et al.
Multiple binding sites for bacterial superantigens on soluble class II MHC molecules
Immunity
(1995)
J.L. Greene et al.
Covalent dimerization of CD28/CTLA-4 and oligomerization of CD80/CD86 regulate T cell costimulatory interactions
J. Biol. Chem
(1996)
H. Tamura et al.
B7-H1 costimulation preferentially enhances CD28-independent T-helper cell function
Blood
(2001)
V.K. Kuchroo et al.
B7-1 and B7-2 costimulatory molecules activate differentially the Th1/Th2 developmental pathways: application to autoimmune disease therapy
Cell
(1995)

D.J. Lenschow et al.

CD28/B7 system of T cell costimulation

Annu. Rev. Immunol

(1996)

E.A. Greenfield et al.

CD28/B7 costimulation: a review

Crit. Rev. Immunol

(1998)

M.L. Alegre et al.

T-cell regulation by CD28 and CTLA-4

Nat. Rev. Immunol

(2001)

B. Salomon et al.

Complexities of CD28/B7: CTLA-4 costimulatory pathways in autoimmunity and transplantation

Annu. Rev. Immunol

(2001)

P.A. van der Merwe et al.

CD80 (B7-1) binds both CD28 and CTLA-4 with a low affinity and very fast kinetics

J. Exp. Med

(1997)

D.A. Ostrov et al.

Structure of murine CTLA-4 and its role in modulating T cell responsiveness

Science

(2000)

C.C. Stamper et al.

Crystal structure of the B7-1/CTLA-4 complex that inhibits human immune responses

Nature

(2001)

J.C. Schwartz et al.

Structural basis for co-stimulation by the human CTLA-4/B7-2 complex

Nature

(2001)

H. Dong et al.

B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion

Nat. Med

(1999)

G.J. Freeman et al.

Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation

J. Exp. Med

(2000)

Cited by (181)

Preclinical development and clinical studies of targeted JAK/STAT combined Anti-PD-1/PD-L1 therapy
2024, International Immunopharmacology
Programmed cell death protein 1 (PD-1) binds to its ligand to help tumours evade the immune system and promote tumour progression. Although anti-PD-1/PD-L1 therapies show powerful effects in some patients, most patients are unable to benefit from this treatment due to treatment resistance. Therefore, it is important to overcome tumour resistance to PD-1/PD-L1 blockade. There is substantial evidence suggesting that the JAK/STAT signalling pathway plays a significant role in PD-1/PD-L1 expression and anti-PD-1/PD-L1 treatment. Herein, we describe the effects of the JAK/STAT signalling pathway on PD-1/PD-L1. Subsequently, the relationship between molecular mutations in the JAK/STAT signalling pathway and immune resistance was analysed. Finally, the latest advancements in drugs targeting the JAK/STAT pathway combined with PD1/PD-L1 inhibitors are summarised.
Review of immune checkpoint blockade and PD-L1 testing in breast cancer
2024, Diagnostic Histopathology
In the past two decades, several drugs have been developed to modulate the immune checkpoints. These modulators regulate, amongst others, the T-cell mediated immune response and may be involved in the escape from immune surveillance. Mounting evidence in several solid tumours points towards the effectiveness of immune checkpoint blockade (ICB) therapy used either as monotherapy or in combination with conventional chemotherapy. In breast cancer, ICB therapy is effective in the treatment of triple negative breast cancer (TNBC). Several PD-L1 companion diagnostic tests have been developed for the selection of patients more likely to benefit from this treatment. Currently, two PD-L1 assays are approved for clinical use in TNBC patients: the SP142 CDx for the use of atezolizumab and the 22C3 PharmDx for the use of pembrolizumab. This review provides the background of PD-L1 testing in breast cancer and discusses the analytical performance of these tests, their scoring algorithms and inter-observer concordance and outlines best practice, including tissue selection, tools for interpretation and use of controls.
Recent approaches for the treatment of uveal melanoma: Opportunities and challenges
2024, Critical Reviews in Oncology/Hematology
Uveal melanoma (UM) is the most prevalent primary intraocular cancer in adult population. Primary methods for treatment of UM involves surgery Proton Beam Therapy (PBT), Plaque Brachytherapy, phototherapy, and Charged Particle Radiation Therapy (CPT). It has been found that approximately 50 % of patients diagnosed with UM ultimately experience development of metastatic disease. Furthermore, it has been identified that majority of the patient experience metastasis in liver with a prevalence of 95 %. Management of metastatic UM (MUM) involves various therapeutic modalities, including systemic chemotherapy, molecular targeted therapy, immunotherapy and liver directed interventions. We outline gene mutation in UM and addresses various treatment modalities, including molecular targeted therapy, miRNA-based therapy, and immunotherapy. Additionally, inclusion of ongoing clinical trials aimed at developing novel therapeutic options for management of UM are also mentioned.
Meta-analysis and Critical Review: Association Between Radio-induced Lymphopenia and Overall Survival in Solid Cancers
2023, Advances in Radiation Oncology
Immune system modulation, with the use of immune checkpoint inhibitors, has drastically changed the field of oncology. Strong preclinical data indicate that radiation therapy (RT) may enhance the response rate to such drugs via in situ vaccination, although these data do not consider immune radiotoxicity. This meta-analysis investigates whether radio-induced lymphopenia (RIL) is associated with overall survival (OS).
A systematic literature search and quantitative analysis were planned, conducted, and reported per the Preferred Reporting Items for Systematic Reviews and Meta-analyses and Quality of Reporting of Meta-analyses checklists. The literature from January 1990 to March 2021 was searched to identify clinical studies with OS data in patients treated with RT and presenting with lymphopenia. A random-effect model was employed for the meta-analysis. Heterogeneity was assessed using the I² statistic. Publication bias was estimated using a P-curve analysis.
A total of 56 studies with 13 223 patients and 11 types of cancers were selected. The mean follow-up time was 35.9 months. Over a third of patients had RIL (37.25%). After removing outlying studies (n = 14), the between-study heterogeneity variance was estimated at t² = 0.018 (P = .01) with an I² value of 36.0% (95% confidence interval, 6%-56%). The results showed that RIL was significantly associated with worse OS (hazard ratio: 1.70; 95% confidence interval, 1.55-1.86; P < .01; 95% prediction interval, 1.27-2.26). A subgroup analysis was performed based on the type of primary tumor, and a difference between the subgroups was found (P < .01). Based on the P-curve analysis, a significant evidential value was found, and no significant publication bias was identified among the studies.
RIL is a significant prognostic factor for mortality in virtually all solid cancers. Pooled-effect estimates indicate a significantly reduced risk of death in patients without RIL. Tailoring RT regimens to spare the immune system and updating dosimetric constraints for new organs at risk, such as major blood vessels, organs with rich blood supplies, bones, and all lymph node areas, may improve prognoses.
Tissue and circulating PD-L2: moving from health and immune-mediated diseases to head and neck oncology
2022, Critical Reviews in Oncology/Hematology
Amongst the chief targets of immune-checkpoint inhibitors (ICIs), namely the Programmed cell death protein 1 (PD-1)/PD-Ligands (Ls) axis, most research has focused on PD-L1, while to date PD-L2 is still under-investigated. However, emerging data support PD-L2 relevant expression in malignancies of the head and neck area, mostly in head and neck squamous cell carcinoma (HNSCC) and salivary gland cancers (SGCs). In this context, ICIs have achieved highly heterogeneous outcomes, emphasizing an urgent need for the identification of predictive biomarkers. With the present review, we aimed at describing PD-L2 biological significance by focusing on its tissue expression, its binding to PD-1 and RGMb receptors, and its impact on physiological and anti-cancer immune response. Specifically, we reported PD-L2 expression rates and significant clinical correlates among different head and neck cancer histotypes. Finally, we described the biology of soluble PD-L2 form and its potential application as a prognostic and/or predictive circulating biomarker.
A novel peptide-based probe <sup>99m</sup>Tc-PEG<inf>6</inf>-RD-PDP2 for the molecular imaging of tumor PD-L2 expression
2022, Chinese Chemical Letters
Tumor-related PD-L2 expression is associated with the clinical efficacy of PD-1/PD-L1 blockade therapy. PD-L2-specific imaging can help selecting patients for appropriate immunotherapy. In this study, a PD-L2-targeting peptide (PDP2) was screened by the one-bead one-compound combinatorial library approach. Using the retro-inverso d-peptide of PDP2 (RD-PDP2) and PEGylation strategies, we developed a novel Tc-99m-labeled PD-L2-targeting peptide as a SPECT tracer (^99mTc-PEG₆-RD-PDP2) for imaging of tumor PD-L2 expression. The radiolabeling yield of ^99mTc-PEG₆-RD-PDP2 was greater than 95% by the standard HYNIC/tricine/TPPTS labeling procedure. ^99mTc-PEG₆-RD-PDP2 displayed high PD-L2-binding specificity both in vitro and in vivo. SPECT/CT imaging with ^99mTc-PEG₆-RD-PDP2 showed that the A549-PD-L2 tumors were clearly visualized, whereas the signals in PD-L2-negative A549 tumors were much lower. In vivo blocking study suggested that the tumor uptake of ^99mTc-PEG₆-RD-PDP2 was PD-L2 specifically mediated. ^99mTc-PEG₆-RD-PDP2 is a promising SPECT probe for the non-invasive imaging of tumor PD-L2 expression and has a great potential in guiding the anti-PD-1 or anti-PD-L1 immunotherapy of cancer.

View all citing articles on Scopus

^☆: Abbreviations: PD-1, programmed death-1; SPR, surface plasmon resonance; MFI, mean fluorescence intensity; RU, response unit; k_on, association rate constant; k_off, dissociation rate constant; R_max, maximum response.

View full text

Differential binding properties of B7-H1 and B7-DC to programmed death-1☆

Abstract

Section snippets

Materials and methods

Characterization of B7-H1Ig, B7-DCIg, and PD-1Ig fusion proteins

Discussion

Acknowledgements

Immunity

Immunity

Immunity

Trends Immunol

Immunity

Biochem. Biophys. Res. Commun

Immunity

J. Biol. Chem

Blood

Cell

CD28/B7 system of T cell costimulation

Annu. Rev. Immunol

CD28/B7 costimulation: a review

Crit. Rev. Immunol

T-cell regulation by CD28 and CTLA-4

Nat. Rev. Immunol

Complexities of CD28/B7: CTLA-4 costimulatory pathways in autoimmunity and transplantation

Annu. Rev. Immunol

CD80 (B7-1) binds both CD28 and CTLA-4 with a low affinity and very fast kinetics

J. Exp. Med

Structure of murine CTLA-4 and its role in modulating T cell responsiveness

Science

Crystal structure of the B7-1/CTLA-4 complex that inhibits human immune responses

Nature

Structural basis for co-stimulation by the human CTLA-4/B7-2 complex

Nature

B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion

Nat. Med

Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation

J. Exp. Med