Compositions and methods for modulating lymphocyte activity

a composition and lymphocyte technology, applied in the field of immunomodulatory compositions and methods, can solve the problems of often insufficient magnitude of immune responses to many different antigens, which are detectable, and cannot afford to protect against disease processes mediated by agents, so as to improve the outcome of organ and tissue transplantation, improve the effect of btla-mediated signaling, and reduce the activity of autoreactive lymphocytes

Inactive Publication Date: 2012-08-23
WASHINGTON UNIV IN SAINT LOUIS
View PDF4 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0017]Also disclosed for the first time herein is the identification of the new B7 family member B7x as a ligand for BTLA, as well as the previously unrecognized ability of B7x to negatively regulate B and T lymphocyte activity through its interaction with BTLA. The interaction of B7x with BTLA inhibits both B and T cell responses, and is a means by which B7x-positive tumor tissue inhibits the activity of tumor-specific T cells. Further, the present disclosure establishes that B7x is also expressed on non-tumor non-lymphoid tissue, thus identifying the B7x / BTLA interaction as a mechanism for maintaining immunological tolerance. These novel findings enable the use of therapeutic agents capable of interfering with the interaction of BTLA and B7x to modulate lymphocyte activity for the purpose of treating, among other conditions, cancer and autoimmune diseases.
[0037]In another preferred embodiment, bioactive agents and methods for mimicking or enhancing the interaction of B7x-positive non-tumor non-lymphoid cells with BTLA-positive T cells are provided, thereby decreasing T cell activity. In an especially preferred embodiment, the T cell is a CD4+ T cell or a CD8+ T cell. In a further embodiment, the CD4+ T cell is a Th1 cell.
[0038]In a further aspect, methods for treating cancers characterized by the presence of B7x-expressing tumor cells are provided. In one embodiment, these methods comprise administering to a mammalian subject at least one of the antagonists of BTLA-mediated signaling disclosed herein, either alone or in conjunction with alternative cancer immunotherapy, chemotherapy and / or radiotherapy protocols. In a preferred embodiment, at least one BTLA or B7x blocking agent is administered to a subject having B7x-positive tumor cells, wherein said blocking agent is capable of interfering with the interaction of BTLA and B7x and inhibiting BTLA-mediated signaling. Preferably, administration of said blocking agents is effective to increase T cell activity directed against tumor antigens other than B7x on the tumor cells, and in particular, to increase cytotoxic T cell activity. Still more preferably, administration of the subject antagonists is effective to inhibit the growth of the B7x-expressing tumor cells.
[0040]In a further aspect, methods for treating autoimmune disorders characterized by the absent or aberrant expression of B7x in non-tumor non-lymphoid host cells subjected to autoimmune attack are provided. In one embodiment, these methods comprise administering to a mammalian subject at least one of the agonists of BTLA-mediated signaling disclosed herein, either alone or in conjunction with alternative immunotherapy and / or immunosuppressive protocols. In a preferred embodiment, at least one BTLA or B7x mimicking agent is administered to a subject having autoreactive BTLA-positive lymphocytes, wherein said mimicking agent is capable of replacing and / or augmenting the interaction of BTLA and B7x and replacing or increasing BTLA-mediated signaling. Preferably, administration of said mimicking agents is effective to decrease autoreactive lymphocyte activity directed against non-tumor non-lymphoid host cells, and particularly autoreactive CD8+ CTL and CD4+ Th1 activity, and B cell activity.
[0041]In a still further aspect, methods for improving the outcome of organ and tissue transplantation and prolonging graft survival are provided. In one embodiment, these methods comprise administering to a transplant recipient at least one of the agonists of BTLA-mediated signaling disclosed herein, either alone or in conjunction with alternative immunotherapy and / or immunosuppressive protocols. In a preferred embodiment, at least one BTLA or B7x mimicking agent is administered to the transplant recipient, wherein said mimicking agent is capable of replacing and / or augmenting the interaction of BTLA and B7x and replacing or increasing BTLA-mediated signaling. Preferably, administration of said mimicking agents is effective to decrease the recipient immune response against donor antigens present in the graft, particularly the cytolytic CTL response and the B cell response. Still more preferably, administration of the subject mimicking agents is effective to bias to T helper cell response from an unfavorable Th-1 type response to a more favorable Th-2 type response, as described in more detail herein.

Problems solved by technology

In addition, immune responses to many different antigens (e.g., microbial antigens or tumor antigens), while detectable, are frequently of insufficient magnitude to afford protection against a disease process mediated by agents (e.g., infectious microorganisms or tumor cells) expressing those antigens.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Compositions and methods for modulating lymphocyte activity
  • Compositions and methods for modulating lymphocyte activity
  • Compositions and methods for modulating lymphocyte activity

Examples

Experimental program
Comparison scheme
Effect test

examples 1-3

Methods for Examples 1-3

[0355]Mice and cells. Female BALB / c were purchased from Jackson Laboratories (Bar Harbor, Me.) and were used at ages 6-9-weeks-old. Animals were housed in accordance with the Animal Care and Use Committee regulations at the University of California, Berkeley. All cell purifications were performed with magnetic cell sorting separation columns (Milternyi Biotec, Auburn, Calif.) with purities >95%. Macrophages were obtained from peritoneal cavity. All cells were cultured in DMEM supplemented with 10% fetal calf serum, 2.mu·M L-glutamine, and 100 U / ml penicillin and streptomycin (all from BioWhittaker), and 2.mu·M 2-mercaptoethanol (Sigma).

[0356]Production of fusion protein. B7xIg was prepared by fusing the coding region of the extracellular domain of B7x to a chimeric sequence containing the CH2-CH3 domain of mouse IgG1 and a Myc-His-tag in pcDNA4 (a gift from Dr. William Sha, UC Berkeley). The construct was linearized with Bgl II and transfected into 293T cells...

example 1

Expression of B7x

[0363]The expression of B7x mRNA in human and mouse tissues was analyzed by Northern blot hybridization. Human B7x was present in a single 3.2-kb mRNA readily detectable in kidney, liver, spleen and placenta. Mouse B7x had three transcripts of 1.9, 3.5 and 8.2 kb, and was expressed significantly in liver, testis, kidney, lung and heart (FIG. 7). With RT-PCR, B7x mRNA was also detected in mouse spleen, prostate, lymph node, thymus, eye, pancreas, B cells, T cells, macrophages, and dendritic cells. (FIG. 8, and data not shown).

[0364]Interestingly, 5 of 8 mouse B7x ESTs located in database searches had been derived from mammary tumors, and 3 of 6 human B7x ESTs originated from ovarian and uterine tumors. To determine whether expression of B7x might be a regular feature of tumors, we used Northern blot analysis to examine a panel of mouse tumors for B7x mRNA expression. Most of the tumor cell lines tested, including NB41A3 (neuroblastoma), P815 (mastocytoma), L1210 (lym...

example 2

B7x Inhibits T Cell Activation Processes

[0367]Initial experiments used purified T cells activated with plate-bound anti-CD3 in the presence of different amounts of immobilized B7xIg. B7xIg decreased proliferation and IL-2 production in a dose-dependent fashion. Additional experiments with purified T cell subsets showed that B7xIg inhibited both CD4 and CD8 T cell responses (FIG. 14). We next employed a conventional costimulation assay. Purified T cells were activated with different amounts of plate-bound anti-CD3 in the presence of CHO transfectants expressing either GFP, B7.2 or B7x. As expected, T cells stimulated in the presence of B7.2 / CHO exhibited enhanced proliferation and cytokine production compared to control GFP / CHO. In contrast, B7x / CHO significantly reduced T cell proliferation and cytokine production (FIG. 15). In order to determine the effect of B7x on T cell activation in the face of costimulation, we also used CHO cells which expressed B7.2 or coexpressed B7.2 and B...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The present invention provides a novel lymphocyte inhibitory receptor termed BTLA which is expressed on both T and B cells, and identifies B7 family member B7x as interacting with BTLA to attenuate lymphocyte activity. Methods and compositions for modulating BTLA-mediated negative signaling and interfering with the interaction of BTLA and B7x for therapeutic, diagnostic and research purposes are also provided.

Description

STATEMENT OF RELATED APPLICATIONS[0001]This application is a continuation of U.S. application Ser. No. 11 / 875,537, filed Oct. 19, 2007 which is a divisional of U.S. application Ser. No. 10 / 600,997, filed Jun. 20, 2003 which claims priority under 35 U.S.C. §119 to U.S. Provisional Patent Application Ser. No. 60 / 390,653, filed Jun. 20, 2002, and to U.S. Provisional Patent Application Ser. No. 60 / 438,593, filed Jan. 6, 2003, the disclosures of which are expressly incorporated herein in their entirety by reference.FIELD OF THE INVENTION[0002]The present invention relates to immunomodulatory compositions and methods, and in particular, to novel lymphocyte regulatory molecules as well as compositions and methods exploiting the same for therapeutic, diagnostic and research purposes.BACKGROUND OF THE INVENTION[0003]Positive and negative costimulatory signals play critical roles in the modulation of T cell activity, and the molecules that mediate these signals have proven to be effective tar...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): C07K16/28C07K14/705C12N15/12
CPCC07K14/70521C07K2319/30C07K2319/00C07K14/70532
Inventor MURPHY, KENNETHWATANABE, NORIHIKOMURPHY, THERESAYANG, JIANFEI
Owner WASHINGTON UNIV IN SAINT LOUIS
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products