[go: up one dir, main page]

WO1999020787A1 - Regulation d'activation de cellules t dependant des cellules porteuses d'antigenes par la chaine alpha 1 de l'haptoglobine - Google Patents

Regulation d'activation de cellules t dependant des cellules porteuses d'antigenes par la chaine alpha 1 de l'haptoglobine Download PDF

Info

Publication number
WO1999020787A1
WO1999020787A1 PCT/US1998/021697 US9821697W WO9920787A1 WO 1999020787 A1 WO1999020787 A1 WO 1999020787A1 US 9821697 W US9821697 W US 9821697W WO 9920787 A1 WO9920787 A1 WO 9920787A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
haptoglobin
chain
alpha
human
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1998/021697
Other languages
English (en)
Inventor
J. Wayne Streilein
Yong Xie
C.-L. Albert Wang
Yanhua Li
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Boston Biomedical Research Institute Inc
Schepens Eye Research Institute Inc
Original Assignee
Boston Biomedical Research Institute Inc
Schepens Eye Research Institute Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Boston Biomedical Research Institute Inc, Schepens Eye Research Institute Inc filed Critical Boston Biomedical Research Institute Inc
Priority to AU10858/99A priority Critical patent/AU1085899A/en
Publication of WO1999020787A1 publication Critical patent/WO1999020787A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
    • G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
    • G01N2333/4701—Details
    • G01N2333/4713—Plasma globulins, lactoglobulin

Definitions

  • LC Epidermal Langerhans cells
  • Langerhans cells acquire a unique functional ability to activate autologous naive T cells, even in the absence of antigens (Schuler et al . , J. Exp. Med. 161:526, 1985; Xie et al . , J. Dermatol . Sci. 2:263-274, 1996). Accumulating evidence indicates that failure to prevent this functional transformation of Langerhans cells in vivo may induce rampant T cell proliferation and trigger skin diseases, such as psoriasis (Demidem et al . , J. Invest. Derm.
  • Known agents such as the corticosteroids and cyclosporin-A, that have the ability to inhibit antigen presentation are non-tissue specific and have effects and toxicities involving many cell types and tissues. Novel agents that could specifically inhibit antigen presentation would be highly desirable.
  • a polypeptide present in normal human blood that interferes with the capacity of antigen presenting cells of the skin (Langerhans cells) to activate naive autologous T cells.
  • This is the first example of a selective inhibitor that acts only on antigen presenting cells and not on other cells of the immune system such as T lymphocytes.
  • This polypeptide has been identified as one of the chains of the molecule haptoglobin, specifically the alpha- 1 chain.
  • Haptoglobin is a well known molecule whose best known biological function is the capture of cell free hemoglobin in order to prevent both iron loss and kidney damage during hemolysis. Haptoglobin is also a positive acute-phase protein. The ability of the alpha-1 chain of haptoglobin to interfere with Langerhans cell function is unprecedented.
  • the invention is directed to pharmaceutical compositions comprising the alpha- 1 chain of haptoglobin (or active portions thereof) , or the entire haptoglobin molecule, and methods for their use to suppress T-cell dependent inflammation in vivo by inhibiting the ability of antigen presenting cells to activate T cells.
  • specific treatable inflammatory skin diseases include psoriasis, mycosis fungoidosis, atopic dermatitis, exfoliative dermatitis and Sezary's Syndrome.
  • a number of autoimmune, T cell dependent inflammations and infectious diseases would be treatable by the methods of the invention.
  • transplantation rejection e.g., Graft Versus Host Disease
  • vaccine development e.g., vaccine development
  • therapy for infectious diseases where antigen presenting cells play a central role in the pathogenesis
  • the methods of the invention can also be used to inhibit Langerhans cell susceptibility to infection with viruses, especially HIV.
  • the invention is directed to pharmaceutical compositions comprising an antagonist to human haptoglobin-haptoglobin receptor binding and to methods for their use in stimulating a T-cell dependent immune response in vivo by stimulating the ability of antigen presenting cells to activate T cells.
  • This treatment would be appropriate in circumstances in which antigen presenting cells, at the T cell activation site, were inappropriately flooded with haptoglobin.
  • Fig. 1 shows the effect of heat treatment on the abilities of human and mouse serum to prevent mouse epidermal LC cells from acquiring in vi tro the capacity to activate naive, syngeneic T cells.
  • BALB/c LC were cultured in medium with or without supplementation with 10% human serum, heated human serum, mouse serum and heated mouse serum, respectively. After 3 days, the LC were harvested and used as stimulators in syngeneic MELR. Six days later, T cell proliferation was assayed as described in Materials and Methods.
  • (*) indicates mean values significantly lower than positive control (B) , P ⁇ 0.001;
  • Fig. 2A shows the abilities of >100 kD and ⁇ 100 kD fractions of heated human serum to prevent epidermal LC cells from acquiring in vi tro the capacity to activate naive, syngeneic T cells.
  • BALB/c epidermal LC were cultured in medium with or without supplementation with 10% unfractionated heated human serum, ⁇ 100 kD fraction and >100 kD fraction of heated human serum, respectively. After 3 days, the LC were harvested and used as stimulators in syngeneic MELR. Six days later, T cell proliferation was assayed as described in Materials and Methods.
  • (*) indicates mean values significantly lower than positive control (B) , P ⁇ 0.001;
  • Fig. 2B shows SDS-PAGE analysis of (a) heated serum fraction that contains protein species greater than (lane 1) or less than (lane 2) 100 kDa separated by ultrafiltration with Centricon-100 tube (Amicon, Beverly, MA) ; (b) protein component of inhibitory fraction (#5 in Fig.2c) purified by HPLC (lane 3) .
  • Lane 4 is the molecular weight standard;
  • Fig. 2C is a profile of the less than 100 kDa fraction of heated human serum analyzed by reversed-phase HPLC. Eight major peaks appeared, and were collected in different fractions. Each fraction after lyophilization was dissolved in water and added to cultures of epidermal LC to test the inhibitory effect. After 3 days LC were used to stimulate syngeneic naive T cells. Among those fractions only fraction
  • Fig. 2D shows the ability of fraction #5 (see Fig. 2c) to prevent LC from acquiring in vi tro the capacity to activate naive, syngeneic T cells.
  • BALB/c epidermal LC were cultured in medium supplemented (or not) with proteins lyophilized from fraction #5 in Fig. 2c and a mixture of proteins lyophilized from the rest of the fractions except for #5. After 3 days, the LC were washed and used as stimulators in syngeneic MELR. Six days later, T cell proliferation was assayed as described in Materials and Methods.
  • (*) indicates mean values significantly lower than positive control (B) , P ⁇ 0.001;
  • Fig. 3 shows the ability of Hp and heated Hp to prevent mouse epidermal LC from acquiring in vi tro the capacity to activate naive T cells, and the effect of heated Hp on T cell proliferation.
  • BALB/c epidermal LC were cultured in medium with or without supplementation with 0.25 mg/ml purified Hp, heated Hp from pooled human serum (Sigma, St. Louis, MO). After 3 days, the LC were washed and used as stimulators in syngeneic MELR. Six days later, T cell proliferation was assayed as described in Materials and Methods.
  • Fig. 4 shows FACS analysis of la and B7-2 expression on freshly obtained mouse LC, and LC cultured in medium supplemented (or not) with .heated Hp .
  • LC were fixed with 5% formaline in PBS and stained with FITC-conjugated mAb anti-mouse I -Ad (Pharmingen, San Diego, CA) followed by PE-conjugated anti-mouse B7-2 (Pharmingen) .
  • FITC-mouse IgG3 and PE-rat IgG2a were used as isotype controls.
  • A freshly obtained LC
  • B LC cultured with medium alone
  • C LC cultured with medium supplemented with heated Hp
  • Fig. 5 shows the effect of Hp-free serum on the ability of human epidermal LC to acquire in vi tro the capacity to activate autologous, naive T cells.
  • Human epidermal LC were cultured in medium supplemented with 20% Hp-free human serum, 20% Hp-free human serum+purified Hp (0.25 mg/ml), or 20% normal human serum. After 3 days, the LC were washed and used as stimulators in human autologous MELR. Six days later, T cell proliferation was assayed as described in
  • Hp-free serum or (C) with 20% Hp-free serum+purified Hp; or
  • Hp-free human serum was obtained by passing serum through immunoaffinity column of anti-human haptoglobin antibody-Sepharose . (Depletion of Hp from serum was confirmed by Western Blotting assay.) ;
  • Fig. 6 is a dose response curve of the inhibitory effect of Hp on LC functional transformation in culture.
  • Human epidermal LC were cultured in medium supplemented with 0.1, 0.2 and 0.4 mg/ml Hp, respectively. After 3 days, the LC were washed and used as stimulators in human autologous MELR. Six days later, T cell proliferation was assayed as described in Materials and Methods.
  • A unstimulated T cells
  • B T cells stimulated with LC that had been cultured in medium alone
  • C T cells stimulated with LC cultured in medium supplemented with 0.4 mg/ml Hp; or
  • D with 0.2 mg/ml Hp; or
  • E with 0.1 mg/ml Hp.
  • (*) indicates mean values significantly lower than control (A), P ⁇ 0.01;
  • Fig. 7 shows confocal laser scanning microscopy of LC in human epidermis stained with anti-HLA and anti-Hp antibodies.
  • Human epidermis were stainded as described in the method and examined with a Leica TCS 4D confocal laser microscope.
  • a dendritic epidermal cell indicated with positive staining of Hp (red) displayed positive staining of HLA (green) in freshly obtained epidermis.
  • the co-localization of Hp and HLA can be viewed in HLA + Hp double staining; and Fig.
  • FIG. 8 shows confocal laser scanning microscopy of LC stained with anti-HLA (green) and anti-Hp (red) antibodies in freshly obtained epidermis (A) , in epidermis cultured for 3 days in Hp-free medium (B) , and in epidermis cultured for 3 days in Hp-containing medium (C) .
  • Epidermal Langerhans cells a subpopulation of dendritic cells, are the primary antigen presenting cells of the skin. As such, they have proven to be functionally plastic. Their precise antigen presenting function is dictated by soluble factors in the microenvironment in which they reside. Epidermal LC carry out two distinct antigen presenting roles in cutaneous immunity. Within the epidermis of normal individuals, LC capture, process and present antigens in a manner that can directly activate previously primed, but not naive, T cells. These are also features of LC freshly prepared from skin and assayed in vi tro .
  • LC When LC are stimulated in vi tro with GM-CSF and IL-1, their functional properties transform in a way that enables the cells to present antigens to naive, unprimed T cells. It is believed that a similar transformation occurs when LC migrate from the skin to draining lymph nodes during the induction of immunity to cutaneous antigens.
  • the ability of cultured LC, but not fresh LC, to activate syngeneic or autologous T cells in the absence of cognate antigen is a convenient method to distinguish the two polar functional phenotypes of LC.
  • Haptoglobin is a well known molecule whose existence and certain functions have been known for almost a hundred years. The best known biological function of haptoglobin is capture of hemoglobin to prevent both iron loss and kidney damage during hemolysis. Haptoglobin is also a positive acute-phase protein and is characterized by a molecular heterogeneity with three major phenotypes: Hp 1-1, Hp 2-2, and the heterozygous Hp 2-1. Although haptoglobin is found in serum of all mammals, this polymorphism exists only in humans. The geographic distribution of haptoglobin phenotypes has been under a strong genetic pressure, and functional differences have been described among the three phenotypes .
  • Haptoglobin polymorphism appears to be related to immune response and to autoimmune and inflammatory disorders.
  • Haptoglobin consists of two different polypeptide chains, the ⁇ -chain and the 3-chain.
  • the ⁇ -chain (40 kD) is heavier than the ⁇ -chain and is identical in all haptoglobin types.
  • Haptoglobin polymorphism arises from variant - chains.
  • the Hp 1-1 phenotype expresses only ⁇ -chains (8.9 kD) .
  • 2 -Chains (16 kD) are present in Hp 2-2 and Hp 2- 1.
  • the higher hemoglobin binding capacity of Hp 1-1 and the association of the Hp-2 allele with higher immune reactivity appear to have contributed to the selection and worldwide distribution of haptoglobin alleles.
  • the haptoglobins display immunoregulative abilities, which can be immunosuppressive by inhibiting lymphocyte reactivity, or immunoinduc ive by influencing IgM biosynthesis adapted to the functional requirements.
  • Hp 2-2 has a stronger effect than the other two haptoglobin phenotypes (Lange, V., Anthropol . Anz . 5_0: 281-302, 1992).
  • Hp 2-2 is associated with higher 5- year mortality of individuals with HIV infection (40% vs. 20% in the Hp 1-1 and 2-1 phenotypes) and is also overrepresented in patients with autoimmune disease. (Langlois et al . , Clin.
  • the human factor was then isolated using a purification protocol consisting of serial procedures of heat treatment, ultrafiltration and high pressure liquid chromatography (HPLC) .
  • HPLC high pressure liquid chromatography
  • human and mouse sera were heated to 85°C for 25 min to determine the heat stability of the factor (s). Heat-denatured proteins precipitated out of the sera following this treatment . The precipitates were removed by centrifugation, and the supernatants were added to culture wells containing fresh murine LC plus GM-CSF.
  • This experiment was designed to determine whether the readily available murine LC could be used as an assay system during the purification procedure. As can be seen in Fig.
  • pooled human serum was then heat treated as described above.
  • the clear supernatant was subjected to ultrafiltration (Centricon-100 filter) in order to separate it into fractions >100kD and ⁇ 100 kD.
  • An aliquot from each fraction was then added to culture wells containing freshly procured murine LC plus GM-CSF. After 3 days, the LC were removed and used to stimulate syngeneic murine T cells.
  • Fig. 2A only the fraction that contained molecules ⁇ 100 kD displayed inhibitory activity. This result was also anomalous compared to the behavior of the factor in whole serum where the inhibitory activity was in the >100kD fraction.
  • fraction no. 5 displayed inhibitory activity (see Fig. 2D) .
  • the inhibition mediated by fraction no. 5 was somewhat less than that of the unfractionated ⁇ 100 kD fraction of heated serum, probably due to losses during the HPLC and lyophilization purification procedures. Susceptability to pronase treatment verified that the inhibitory factor was proteinaceous .
  • N-terminus sequencing of up to 20 residues was carried out on an automated gas phase sequenator (ABI 477A) .
  • the sequence was found to be:
  • Unheated human Hp displayed no inhibitory activity.
  • the inhibition of LC effected by heated Hp is not nonspecific, however, because direct addition of heated human Hp to syngeneic MELR failed to inhibit T cell proliferation (Fig. 3, bar E) .
  • the inhibition is also reversible, as cells incubated for 24 h in heated Hp followed by 48 h in Hp-free medium proved to be vigorous activators of syngeneic T cells. This result also suggests that Hp is not toxic to LC.
  • the capacity of Hp to inhibit LC from acquiring the capacity to activate naive, syngeneic T cells may be related to effects on expression of MHC molecules and co-stimulatory molecules. To examine this possibility, murine LC were cultured with heated human Hp .
  • Control LC were cultured in the absence of heated human haptoglobin. After three days, the cells were removed and analyzed by flow cytometry for expression of class II and B7-2 molecules. As shown in Fig. 4, LC cultured in the presence of heated human Hp expressed reduced class II molecules and reduced B7-2 compared to LC cultured in the absence of Hp.
  • Hp is the factor in human serum that inhibits human LC from acquiring the capacity to activate autologous naive T cells in culture.
  • Human serum was passed through an anti-Hp antibody coupled immunoaffinity column in order to deplete the serum of Hp (deletion of Hp was confirmed by Western blot analysis) .
  • Hp-free serum was then added to medium in which fresh human LC were cultured. Three days later, the LC were removed and used to stimulate autologous T cells. The results presented in Fig. 5 show that LC cultured in the presence of Hp-free serum readily activated autologous T cells.
  • anti-human Hp monoclonal antibody was used to stain freshly procured human epidermal sheets. Numerous epidermal cells with dendritic shape were observed with intense positive staining of Hp . When counter-stained with anti-HLA class II antibodies (which stains only LC in normal epidermis) , all Hp-positive cells were confirmed to be LC
  • mice Female BALB/c and C57BL/6 mice were obtained from
  • epidermal LC Preparation of epidermal LC .
  • skin blisters were raised on healthy volunteers after a Dermovac (Turko, Finland) instrument was placed on abdominal skin with negative pressure (about 250 mm Hg) for 2-3 h.
  • the domes of skin blisters were removed using sterile surgical procedures, and floated on cultured medium for 2-3 day culture.
  • Cultured epidermal sheets were digested with 0.25% trypsin at 37°C for 15 min, disaggregated using a 5 ml syringe, and filtered through a 70 ⁇ m cell strainer (Falcon, Franklin Lakes, NJ) . Recovered cells were used as cultured LC.
  • mice LC For mouse LC: shaved mouse skin was removed; subcutaneous tissues was scraped off and the skin sheet was floated on 0.25% trypsin-PBS for 1 h at 37°C. The epidermis was then separated with a fine forceps and incubated in 0.25 trypsin for an additional 10 min. The epidermis was further disaggregated using 10 ml syringe, filtered through 70 ⁇ m cell strainer, and the cell suspension used as LC.
  • T lymphocytes Preparation of highly purified T lymphocytes. Axillary, inguinal and mesenteric lymph nodes were removed from naive mice, minced and resuspended in 2 ml T cell -enrichment buffer
  • the cell suspension was loaded on mouse T cell -enrichment columns (R&D Systems) , incubated at room temperature for 10 min, and then eluted with washing buffer. The first 6 ml of eluted cells were collected and referred to as purified naive T cells. More than 95% of these recovered cells were CD3 positive on the basis of flow cytometry analysis. Human T lymphocytes were purified in the same way from peripheral blood.
  • Immunofluorescence assays Staining of human epidermal sheets: human epidermal sheets were fixed in acetone for 10 min, incubated with mouse anti-human haptoglobin monoclonal antibody (IgGl, 1:80, Sigma) for 60 min at 37°C and rinsed. Cy3-conjugated goat anti-mouse IgGl immunoglobin (1:80, Caltag Laboratories, Burlingame, CA) was applied for 60 min in order to label haptoglobin. FITC-conjugated mouse anti-human HLA-DR, DP, DQ (IgG2a, 1:20, Pharmingen, San Diego, CA) was applied for 60 min to counter-stain for HLA.
  • mouse anti-human haptoglobin monoclonal antibody IgGl, 1:80, Sigma
  • Cy3-conjugated goat anti-mouse IgGl immunoglobin (1:80, Caltag Laboratories, Burlingame, CA) was applied for 60 min
  • Controls consisted of substitution of primary anti-human haptoglobin antibodies with isotype mouse IgGl (Caltag) , and FITC-conjugated isotype mouse IgG2a (Pharmingen) .
  • FACS analysis of la, and B7-2 expression on mouse LC were stained with FITC-conjugated mAb anti-mouse I-Ad (Pharmingen) followed by PE-conjugated anti-mouse B7-2 (Pharmingen) .
  • FITC-mouse IgG3 and PE-rat IgG2a (Pharmingen) were used as isotype controls.
  • Cells were analyzed on a COULTER EPICS XL flow cytometer using Workstation software Version 1.5 (Coulter, FL) .
  • MLR Mixed epidermal cell lymphocyte reactions
  • Purified naive lymph node T cells (3xl0 5 ) were cultured with 5xl0 4 stimulatory cells (fresh LC, cultured LC) in round bottom microtiter plates at 37°C, 5% C0 2 for 6 days.
  • 5xl0 4 stimulatory cells fresh LC, cultured LC
  • One ⁇ Ci 3 H-thymidine was added to each well 16-18 h prior to termination of culture.
  • the cells were harvested with Tomtec Harvester (Wallac, LKB, NJ) and mean ( ⁇ standard error) radioisotope incorporation of 6 replicated wells were calculated by an automatic ⁇ counter (Wallac) .
  • BALB/c LC and BALB/c T cells were used to evaluate the syngeneic stimulatory reaction.
  • BALB/c LC and C57BL/6 T cells were used to evaluate the allogeneic stimulatory reaction.
  • autologous purified peripheral blood T cells and epidermal LC were used. Experiments were repeated at least three times m mice and two times in humans .
  • SDS-PAGE Samples were loaded onto 14% SDS-polyacrylamide gels and run under denaturing condition in the presence of 0.1% SDS and 2.5 mM dithiothreitol . Amino acid sequencing was performed using the pulsed liquid sequencer model 477A (Applied Biosystems, Foster City, CA) . The derived sequence was entered using sequence analyzer software Swiss Prot .
  • LC Langerhans cells
  • epidermal LC are incapable of activating naive, antigen-specific T cells in si tu . It is only when the cells undergo functional transformation, especially in response to GM-CSF, that they acquire the novel capacity to present new antigens to naive, specific T cells. And they do so in draining lymph nodes - not in the skin. It is in respect to these two polar expressions of LC function that our finding of a regulatory role for Hp takes on meaning. Our results indicate that LC resident within the normal epidermis contain Hp, particularly in cytoplasmic vesicles.
  • haptoglobin to inhibit functional changes in LC, specifically, to inhibit the ability of antigen presenting cells to activate T cells by interfering with the presentation of immunogenic peptides on class II MHC molecules and by interfering with up-regulation of co- stimulatory molecules, is expected to extend to other dendritic cells, the leukocyte lineage to which LC belong. Even dendritic cells from peripheral organs have been reported to undergo a functional transformation in vi tro when cultured in the presence of GM-CSF. After this transformation, the cells display potent co-stimulatory features similar to those described for cultured LC (Girolomoni et al . , J. Immunol. 145 :2820, 1990; Dai et al . , Regional Immunology 5_:269, 1993) .
  • the alpha- 1 chain of haptoglobin or the active portion thereof (which can be determined by conventional methods using fragments of the haptoglobin alpha- 1 chain in the assay systems described herein) can be provided as an isolated polypeptide (or stabilized as part of a fusion protein or complexed with the beta chain as the haptoglobin molecule) and used directly in a therapeutic composition to inhibit Langerhans cells and other dendritic cell functional properties and, therefore, can be used to interfere with Langerhans cells/dendritic cell-dependent activation of T cells.
  • T cells There are many human disorders where inappropriate activation of T cells is a major pathogenic parameter.
  • specific treatable inflammatory skin diseases include psoriasis, mycosis fungoidosis, atopic dermatitis, exfoliative dermatitis and Sezary's Syndrome.
  • autoimmune, T cell dependent inflammations, and infectious diseases that would be treatable by the methods of the invention.
  • Specific examples include prevention of transplantation rejection (e.g., Graft Versus Host Disease) , vaccine development, and therapy for infectious diseases where antigen presenting cells play a central role in the pathogenesis .
  • transplantation rejection e.g., Graft Versus Host Disease
  • vaccine development e.g., vaccine development
  • therapy for infectious diseases where antigen presenting cells play a central role in the pathogenesis
  • the methods of the invention could be used to inhibit Langerhans cell susceptibility to infection with viruses, especially HIV.
  • peptide inhibitors i.e., antagonists
  • peptide inhibitors that bind to the haptoglobin receptor or to the haptoglobin alpha-1 chain and inhibit receptor triggering, or anti- haptoglobin alpha-1 chain antibodies
  • This treatment would be most effective in circumstances in which antigen presenting cells, at the T cell activation site, were inappropriately flooded with haptoglobin.
  • the polymorphism of the haptoglobin alpha chain locus would be a useful indicator to diagnose individuals potentially susceptible to diseases resulting from excessive antigen presenting cell stimulation of T cells.
  • compositions containing human haptoglobin, its alpha-1 chain (or the active portion thereof) or inhibitors of haptoglobin receptor binding may be administered, e.g., by injection (include subcutaneous, intravenous and intramuscular) ; by oral and inhalation or aerosol therapy; by application to the skin in a lotion, cream, ointment or paste; or by rectal or vaginal suppositories (more suitable for the prevention of AIDS) .
  • the active compound can be administered in a dosage of, e . g. , about 2 mg/kg/day to 10 mg/kg/day.
  • Optional dosage and modes of administration can readily be determined by conventional protocols.
  • compositions in the methods of the invention may be administered independently or co- administered with another immunosuppressive agent, e.g., cyclosporin-A or corticosteroids .
  • another immunosuppressive agent e.g., cyclosporin-A or corticosteroids .

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Microbiology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne un polypeptide présent dans le sang humain normal et interférant avec la capacité des cellules porteuses d'antigènes de l'épiderme (cellules de Langerhans) d'activer des cellules T autologues naïves, ce polypeptide ayant été découvert et identifié comme étant la chaîne alpha 1 de la molécule d'haptoglobine. La chaîne alpha 1 de l'haptoglobine est un inhibiteur sélectif agissant uniquement sur les cellules porteuses d'antigènes et non sur d'autres cellules du système immunitaire telles que les lymphocytes T. On peut administrer des compositions thérapeutiques contenant de l'haptoglobine humaine ou sa chaîne alpha 1 à un patient, afin d'interférer avec une activation de cellules T dépendant de cellules de Langerhans/cellules dendritiques et, ainsi, supprimer une inflammation in vivo dépendant de cellules T. Les dermatoses inflammatoires spécifiques pouvant être traitées comprennent le psoriasis, le mycosis fongoïde, l'eczéma atopique, la dermatite exfoliatrice et le syndrome de Sézary. On pourrait traiter, grâce aux méthodes selon l'invention, d'autres inflammations ou maladies infectieuses auto-immunes et dépendant de cellules T; on pourrait également prévenir le rejet de greffes, (par exemple la réaction du greffon contre l'hôte), développer des vaccins et un traitement de maladies infectieuses dans la pathogenèse desquelles les cellules porteuses d'antigènes jouent un rôle central.
PCT/US1998/021697 1997-10-17 1998-10-15 Regulation d'activation de cellules t dependant des cellules porteuses d'antigenes par la chaine alpha 1 de l'haptoglobine Ceased WO1999020787A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU10858/99A AU1085899A (en) 1997-10-17 1998-10-15 Control of apc-dependent t cell activation by the alpha-1 chain of haptoglobi

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US6251997P 1997-10-17 1997-10-17
US60/062,519 1997-10-17

Publications (1)

Publication Number Publication Date
WO1999020787A1 true WO1999020787A1 (fr) 1999-04-29

Family

ID=22043008

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1998/021697 Ceased WO1999020787A1 (fr) 1997-10-17 1998-10-15 Regulation d'activation de cellules t dependant des cellules porteuses d'antigenes par la chaine alpha 1 de l'haptoglobine

Country Status (2)

Country Link
AU (1) AU1085899A (fr)
WO (1) WO1999020787A1 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005121328A3 (fr) * 2004-06-11 2006-04-06 Syngenta Ltd Methode permettant d'ameliorer un etat inflammatoire de la peau
US9534029B2 (en) 2012-10-03 2017-01-03 Csl Behring Ag Method of purifying proteins

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
EL GHMATI S. M., ET AL.: "IDENTIFICATION OF HAPTOGLOBIN AS AN ALTERNATIVE LIGAND FOR CD11B/ CD18.", THE JOURNAL OF IMMUNOLOGY, THE AMERICAN ASSOCIATION OF IMMUNOLOGISTS, US, vol. 156., 1 June 1996 (1996-06-01), US, pages 2542 - 2552., XP002915564, ISSN: 0022-1767 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005121328A3 (fr) * 2004-06-11 2006-04-06 Syngenta Ltd Methode permettant d'ameliorer un etat inflammatoire de la peau
WO2005121329A3 (fr) * 2004-06-11 2006-04-27 Syngenta Ltd Procede permettant d'ameliorer un etat inflammatoire de la peau
US9534029B2 (en) 2012-10-03 2017-01-03 Csl Behring Ag Method of purifying proteins
US9937229B2 (en) 2012-10-03 2018-04-10 Csl Behring Ag Methods of treatment using hemopexin compositions
US10918696B2 (en) 2012-10-03 2021-02-16 Csl Behring Ag Methods of treatment using hemopexin compositions

Also Published As

Publication number Publication date
AU1085899A (en) 1999-05-10

Similar Documents

Publication Publication Date Title
Xie et al. Haptoglobin is a natural regulator of Langerhans cell function in the skin
LaSalle et al. Early signaling defects in human T cells anergized by T cell presentation of autoantigen.
Puré et al. Antigen processing by epidermal Langerhans cells correlates with the level of biosynthesis of major histocompatibility complex class II molecules and expression of invariant chain.
Hopfer et al. The importance of cell‐mediated immunity in the course and severity of autoimmune anti‐glomerular basement membrane disease in mice
Omary et al. Human homologue of murine T200 glycoprotein.
Semana et al. T cell autoreactivity to proinsulin epitopes in diabetic patients and healthy subjects
EP1025849B9 (fr) Procedes de detection et de traitement de personnes presentant des cellules anormales exprimant les antigenes peptidiques hla-a2/tyrosinase
CN100482685C (zh) 来源于髓鞘碱性蛋白的耐受原性肽
CA2429579A1 (fr) Cd200 tronquee
AU2013207489A1 (en) Partial MHC constructs and methods of use
CA2110055C (fr) Utilisation de peptides du recepteur de lymphocytes t contre les maladies du systeme immunitaire
EP0463080A1 (fr) Produits pharmaceutiques a base de cd8
Tamburrini et al. Immunosuppressive activity of bovine seminal RNase on T‐cell proliferation
WO2010028066A2 (fr) Épitopes cd133
EP0687180B1 (fr) Procedes de detection et de traitement de personnes presentant des cellules anormales exprimant les antigenes peptidiques hla-a2/tyrosinase
WO2003029293A2 (fr) Modulateurs
La Flamme et al. Trypanosoma cruzi‐infected macrophages are defective in major histocompatibility complex class II antigen presentation
US5530096A (en) Isolated, tyrosinase derived peptides and uses thereof
EP2436394A1 (fr) Thérapie par peptides
EP1384781B1 (fr) Procédés d'inhibition de la survie des cellules T, par inhibition du taux de protéine BCL-XL
Hansen et al. Leukemic T cells from patients with cutaneous T-cell lymphoma demonstrate enhanced activation through CDw60, CD2, and CD28 relative to activation through the T-cell antigen receptor complex
WO1999020787A1 (fr) Regulation d'activation de cellules t dependant des cellules porteuses d'antigenes par la chaine alpha 1 de l'haptoglobine
KR20150105365A (ko) 펩타이드
JP2010235607A (ja) 自己免疫疾患の診断薬と治療薬となるペプチド類
Puré et al. Antigen processing by epidermal Langerhans cells correlates with the level of biosynthesis of MHC class II molecules and expression of invariant chain

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA