WO2012054755A1 - Chlamydia antigens and uses thereof - Google Patents
Chlamydia antigens and uses thereof Download PDFInfo
- Publication number
- WO2012054755A1 WO2012054755A1 PCT/US2011/057143 US2011057143W WO2012054755A1 WO 2012054755 A1 WO2012054755 A1 WO 2012054755A1 US 2011057143 W US2011057143 W US 2011057143W WO 2012054755 A1 WO2012054755 A1 WO 2012054755A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- polypeptide antigen
- antigen
- polypeptide
- chlamydia
- composition
- Prior art date
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/118—Chlamydiaceae, e.g. Chlamydia trachomatis or Chlamydia psittaci
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55561—CpG containing adjuvants; Oligonucleotide containing adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55577—Saponins; Quil A; QS21; ISCOMS
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/572—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 cytotoxic response
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
Definitions
- Chlamydia trachomatis is an obligate intracellular bacterium which exists as multiple serovariants with distinct tropism for the eye or urogenital tract. Infection with urogenital variants can cause various disease conditions such as urethritis, cervicitis, pharyngitis, proctitis, epididymitis, and prostatitis. Untreated chlamydial infection can cause pelvic inflammatory disease, which in turn can lead to ectopic pregnancy, infertility, and chronic pelvic pain. Infection during pregnancy has been linked to severe complications such as spontaneous abortion, premature delivery, premature rupture of fetal membranes, low birth weight, and neonatal infections (Navarro et al., Can. J. Inf. Dis. 13(3): 195-207, 2002).
- C. trachomatis Infection with ocular variants of C. trachomatis can cause trachoma, or conjunctivitis of eyelid and corneal surfaces, and is a leading cause of preventable blindness.
- Pathological effects of C. trachomatis in humans are a significant societal economic burden as well as an ongoing public health concern in both industrialized and developing nations. An estimated four to five million new cases of chlamydial infection occur each year in the United States alone. The annual costs of treating pelvic inflammatory disease may be as high as US $10 billion.
- the prevalence of C. trachomatis infection in the developing world is over 90%, with an estimated 500 million people at high risk for infection (World Health Organization, Sexually Transmitted Diseases, 2008). There is an urgent need for immunogenic, effective vaccines for controlling chlamydial infections worldwide.
- the present invention encompasses the discovery of novel antigens from
- Chlamydia trachomatis that elicit antigen specific immune responses in mammals.
- Such novel antigens, and/or nucleic acids encoding the antigens can be incorporated into immunogenic compositions and administered to elicit immune responses, e.g., to provide protection against chlamydia infections and disease caused by chlamydia organisms.
- Such novel antigens, and/or responses to novel antigens can be detected to identify and/or characterize immune responses to chlamydia organisms.
- the invention provides immunogenic compositions
- chlamydia antigen selected from a CT062 polypeptide antigen, a CT572 polypeptide antigen, a CT043 polypeptide antigen, a CT570 polypeptide antigen, a CT177 polypeptide antigen, a CT725 polypeptide antigen, a CT067 polypeptide antigen, a CT476 polypeptide antigen, and combinations thereof.
- a chlamydia antigen comprises a full-length chlamydia polypeptide.
- a chlamydia antigen comprises a portion or portions of a full-length chlamydia polypeptide.
- a chlamydia antigen comprises a chlamydia polypeptide that lacks a signal sequence and/or trans-membrane domain.
- a chlamydia antigen comprises a mixture of full-length chlamydia polypeptide and fragments resulting from processing, or partial processing, of a signal sequence by an expression host, e.g., E. coli, an insect cell line (e.g.. the baculovirus expression system), or a mammalian (e.g., human or Chinese Hamster Ovary) cell line.
- an expression host e.g., E. coli, an insect cell line (e.g.. the baculovirus expression system), or a mammalian (e.g., human or Chinese Hamster Ovary) cell line.
- an expression host e.g., E. coli, an insect cell line (e.g.. the baculovirus expression system), or a mammalian (e
- an immunogenic composition comprises a CT062 polypeptide antigen.
- a CT062 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of a CT062 polypeptide sequence.
- a CT062 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l.
- a CT062 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l.
- an immunogenic composition comprises a CT572 polypeptide antigen.
- a CT572 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of a CT572 polypeptide sequence.
- a CT572 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3.
- a CT572 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3.
- an immunogenic composition comprises a CT043 polypeptide antigen.
- a CT043 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of a CT043 polypeptide sequence.
- a CT043 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5.
- a CT043 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5.
- an immunogenic composition comprises a CT570 polypeptide antigen.
- a CT570 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of a CT570 polypeptide sequence.
- a CT570 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7.
- a CT570 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7.
- an immunogenic composition comprises a CT177 polypeptide antigen.
- a CT177 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of a CT177 polypeptide sequence.
- a CT177 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9.
- a CT177 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9.
- an immunogenic composition comprises a CT725 polypeptide antigen.
- a CT725 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of a CT725 polypeptide sequence.
- a CT725 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of the sequence shown in SEQ ID NO: 11.
- a CT725 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of the sequence shown in SEQ ID NO: 11.
- an immunogenic composition comprises a CT067 polypeptide antigen.
- a CT067 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of a CT067 polypeptide sequence.
- a CT067 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of the sequence shown in SEQ ID NO:23.
- a CT067 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of the sequence shown in SEQ ID NO:23.
- an immunogenic composition comprises a CT476 polypeptide antigen.
- a CT476 polypeptide antigen comprises at least
- a CT476 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of a CT476 polypeptide sequence.
- a CT476 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of the sequence shown in SEQ ID
- a CT476 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of the sequence shown in SEQ ID NO:63.
- an immunogenic composition comprises a p6 polypeptide antigen from the cryptic plasmid of chlamydia.
- a p6 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of a p6 polypeptide sequence.
- a p6 polypeptide antigen comprises at least 7,
- a p6 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8,
- an immunogenic composition comprises a CT310 polypeptide antigen.
- a CT310 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of a CT310 polypeptide sequence.
- a CT310 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:67.
- a CT310 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:67.
- an immunogenic composition comprises a CT638 polypeptide antigen.
- a CT638 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of a CT638 polypeptide sequence.
- a CT638 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69.
- a CT638 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69.
- an immunogenic composition comprises two or more isolated chlamydia antigens.
- the two or more isolated chlamydia antigens comprise two or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise three or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise four or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise five, six, seven or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise eight polypeptide antigens selected from Table 1.
- an immunogenic composition comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1 ; and (b) one or more chlamydia polypeptide antigens selected from Table 2.
- an immunogenic composition comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1 ; and (b) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 2; and (b) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises three or more isolated chlamydia antigens, wherein the three or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1 ; (b) one or more chlamydia polypeptide antigens selected from Table 2; and (c) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises an isolated chlamydia polypeptide antigen selected from Table 2.
- an immunogenic composition comprises an isolated chlamydia polypeptide antigen selected from Table 3.
- an immunogenic composition comprises two, three, four, five or more isolated chlamydia polypeptide antigens selected from Table 2. [0021] In some embodiments, an immunogenic composition comprises two, three, four, five or more isolated chlamydia polypeptide antigens selected from Table 3.
- a chlamydia antigen is fused to a heterologous polypeptide (e.g., an epitope tag).
- a heterologous polypeptide e.g., an epitope tag
- an immunogenic composition comprising a chlamydia antigen includes a pharmaceutically acceptable excipient.
- an immunogenic composition comprising a chlamydia antigen includes an adjuvant.
- an immunogenic composition includes a mineral-containing adjuvant.
- the mineral-containing adjuvant includes aluminum hydroxide.
- an immunogenic composition includes an adjuvant comprising an immunomodulatory oligonucleotide.
- an immunogenic composition includes IC31TM adjuvant (Intercell AG).
- an immunogenic composition includes an adjuvant comprising a toxin.
- an immunogenic composition includes an adjuvant comprising an endotoxin. In some embodiments, an immunogenic composition includes an adjuvant comprising a muramyl dipeptide. In some embodiments, an immunogenic composition includes an adjuvant comprising an oil emulsion. In some embodiments, an immunogenic composition includes an adjuvant comprising a saponin. In some embodiments, an immunogenic composition includes an adjuvant comprising an immune stimulating complex (ISCOM). In some embodiments, an immunogenic composition includes an adjuvant comprising a nonionic block copolymer. In some embodiments, an immunogenic composition includes virus-like particles (VLPs). In some embodiments, an immunogenic composition includes replicons.
- VLPs virus-like particles
- an immunogenic composition includes an adjuvant comprising lipososmes. In some embodiments, an immunogenic composition includes an adjuvant comprising microparticles. In some embodiments, an immunogenic composition includes an adjuvant comprising biodegradable microspheres. In some embodiments, an immunogenic composition includes an adjuvant comprising a cytokine. In some
- an immunogenic composition includes an adjuvant comprising a lipopeptide.
- an immunogenic composition elicits an immune response to Chlamydia trachomatis.
- an immunogenic composition elicits a T cell-mediated immune response to a chlamydia antigen (e.g., a CD4 + T cell- mediated immune response and/or a CD8 + T cell-mediated immune response).
- a chlamydia antigen e.g., a CD4 + T cell- mediated immune response and/or a CD8 + T cell-mediated immune response.
- an immunogenic composition elicits a Thl T cell response.
- an immunogenic composition elicits a Thl7 T cell response.
- an immunogenic composition elicits IFN- ⁇ secretion by antigen-specific T cells.
- an immunogenic composition elicits a cytotoxic T cell response.
- an immunogenic composition elicits an antibody response (e.g., an IgG response, and/or an IgA response). In some embodiments, an immunogenic composition elicits a B cell-mediated immune response. In some embodiments, an immunogenic composition elicits both a T cell- and a B cell-mediated response. In some embodiments, an immunogenic composition elicits an innate immune response.
- an antibody response e.g., an IgG response, and/or an IgA response
- an immunogenic composition elicits a B cell-mediated immune response.
- an immunogenic composition elicits both a T cell- and a B cell-mediated response. In some embodiments, an immunogenic composition elicits an innate immune response.
- the invention provides methods for eliciting an immune response against chlamydia in a mammal.
- the methods include, for example, administering to the mammal an immunogenic composition comprising an isolated chlamydia polypeptide antigen selected from Table 1, Table 2, or Table 3, or combinations thereof, e.g., an immunogenic composition described herein.
- a method elicits an immune response against
- a method elicits a T cell response to a chlamydia antigen (e.g., a CD4 + T cell mediated immune response and/or a CD8 + T cell mediated immune response).
- a method elicits a Thl T cell response.
- a method elicits a Thl7 T cell response.
- a method elicits IFN- ⁇ secretion by antigen-specific T cells.
- a method elicits an antibody response (e.g., an IgG response, and/or an IgA response).
- a method elicits a cytotoxic T cell response. In some embodiments, a method elicits a B cell-mediated immune response. In some embodiments, a method elicits both a T cell- and a B cell-mediated response. In some embodiments, a method elicits an innate immune response.
- a method reduces the incidence of chlamydia infection in subjects administered the composition. In some embodiments, a method reduces the likelihood of lower tract infection by a chlamydia organism. In some embodiments, a method reduces the likelihood of upper tract infection by a chlamydia organism. In some
- a method reduces the likelihood of chronic infection by a chlamydia organism. In some embodiments, a method reduces the likelihood of suffering from pelvic inflammatory disease due to a chlamydia infection. In some embodiments, a method reduces the likelihood of infertility subsequent to a chlamydia infection.
- an immunogenic composition is administered to the mammal at least two times (e.g., two, three, four, or five times).
- an immunogenic composition administered after a first administration differs from the composition administered initially, e.g., the composition includes a different chlamydia antigen or a different subset of chlamydia antigens, or a different chlamydia antigen substance (polypeptide or nucleic acid encoding same), or a different dose of antigen, or a different adjuvant, or a different dose of adjuvant.
- a boost is administered by a different route than a previous administration.
- the mammal is at risk for infection with Chlamydia trachomatis. In some embodiments, the mammal is infected with Chlamydia trachomatis. In some embodiments, the mammal is a female. In some embodiments, the mammal is a human.
- an immunogenic composition administered in a method comprises an adjuvant.
- an adjuvant is a mineral-containing adjuvant.
- an immunogenic composition administered in a method comprises a pharmaceutically acceptable excipient.
- an immunogenic composition comprises an adjuvant.
- an immunogenic composition includes a mineral-containing adjuvant.
- a mineral-containing adjuvant includes aluminum hydroxide.
- an immunogenic composition includes an adjuvant comprising an
- an immunogenic composition includes IC31TM adjuvant (Intercell AG). In some embodiments, an immunogenic composition includes an adjuvant comprising a toxin. In some embodiments, an
- an immunogenic composition includes an adjuvant comprising an endotoxin. In some embodiments, an immunogenic composition includes an adjuvant comprising a muramyl dipeptide. In some embodiments, an immunogenic composition includes an adjuvant comprising an oil emulsion. In some embodiments, an immunogenic composition includes an adjuvant comprising a saponin. In some embodiments, an immunogenic composition includes an adjuvant comprising an immune stimulating complex (ISCOM). In some embodiments, an immunogenic composition includes an adjuvant comprising a nonionic block copolymer. In some embodiments, an immunogenic composition includes virus-like particles (VLPs). In some embodiments, an immunogenic composition includes replicons.
- VLPs virus-like particles
- an immunogenic composition includes an adjuvant comprising lipososmes. In some embodiments, an immunogenic composition includes an adjuvant comprising microp articles. In some embodiments, an immunogenic composition includes an adjuvant comprising biodegradable microspheres. In some embodiments, an immunogenic composition includes an adjuvant comprising a cytokine. In some embodiments, an immunogenic composition includes an adjuvant comprising a lipopeptide.
- an immunogenic composition comprises a CT062 polypeptide antigen.
- a CT062 polypeptide antigen comprises 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of a CT062 polypeptide sequence.
- a CT062 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l.
- a CT062 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l.
- an immunogenic composition comprises a CT572 polypeptide antigen.
- a CT572 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of a CT572 polypeptide sequence.
- a CT572 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3.
- a CT572 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3.
- an immunogenic composition comprises a CT043 polypeptide antigen.
- a CT043 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of a CT043 polypeptide sequence.
- a CT043 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5.
- a CT043 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5.
- an immunogenic composition comprises a CT570 polypeptide antigen.
- a CT570 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of a CT570 polypeptide sequence.
- a CT570 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7.
- a CT570 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7.
- an immunogenic composition comprises a CT177 polypeptide antigen.
- a CT177 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of a CT177 polypeptide sequence.
- a CT177 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9.
- a CT177 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9.
- an immunogenic composition comprises a CT725 polypeptide antigen.
- a CT725 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of a CT725 polypeptide sequence.
- a CT725 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of the sequence shown in SEQ ID NO: 11.
- a CT725 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of the sequence shown in SEQ ID
- an immunogenic composition comprises a CT067 polypeptide antigen.
- a CT067 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of a CT067 polypeptide sequence.
- a CT067 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of the sequence shown in SEQ ID NO:23.
- a CT067 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of the sequence shown in SEQ ID NO:23.
- an immunogenic composition comprises a CT476 polypeptide antigen.
- a CT476 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of a CT476 polypeptide sequence.
- a CT476 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of the sequence shown in SEQ ID NO:63.
- a CT476 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of the sequence shown in SEQ ID NO:63.
- an immunogenic composition comprises a p6 polypeptide antigen from the cryptic plasmid of chlamydia.
- a p6 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of a p6 polypeptide sequence.
- a p6 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of the sequence shown in SEQ ID NO:65.
- a p6 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of the sequence shown in SEQ ID NO:65.
- an immunogenic composition comprises a CT310 polypeptide antigen.
- a CT310 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of a CT310 polypeptide sequence.
- a CT310 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:67.
- a CT310 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:67.
- an immunogenic composition comprises a CT638 polypeptide antigen.
- a CT638 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of a CT638 polypeptide sequence.
- a CT638 polypeptide antigen comprises at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69.
- a CT638 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69.
- an immunogenic composition comprises two or more isolated chlamydia antigens.
- the two or more isolated chlamydia antigens comprise two or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise three or more of a polypeptide antigen selected from Tablel.
- the two or more isolated chlamydia antigens comprise four or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise five, six, seven or more of a polypeptide antigen selected from Table 1.
- the two or more isolated chlamydia antigens comprise eight polypeptide antigens selected from Table 1.
- an immunogenic composition suitable for a method of the invention comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1; and (b) one or more chlamydia polypeptide antigens selected from Table 2.
- an immunogenic composition comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1 ; and (b) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises two or more isolated chlamydia antigens, wherein the two or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 2; and (b) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises three or more isolated chlamydia antigens, wherein the three or more isolated chlamydia antigens comprise (a) one or more chlamydia polypeptide antigens selected from Table 1 ; (b) one or more chlamydia polypeptide antigens selected from Table 2; and (c) one or more chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises an isolated chlamydia polypeptide antigen selected from Table 2.
- an immunogenic composition comprises an isolated chlamydia polypeptide antigen selected from Table 3.
- an immunogenic composition comprises two, three, four, five or more isolated chlamydia polypeptide antigens selected from Table 2.
- an immunogenic composition comprises two, three, four, five or more isolated chlamydia polypeptide antigens selected from Table 3.
- an immunogenic composition comprises a chlamydia antigen and an antigen from a different infectious agent.
- an immunogenic composition comprises a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof; and an antigen from a papillomavirus (e.g., a human papillomavirus).
- an immunogenic composition comprises a chlamydia polypeptide antigen selected from Table 1 , Table 2, Table 3, or combinations thereof; and an antigen from a herpesvirus (e.g., herpes simplex virus-2).
- a herpesvirus e.g., herpes simplex virus-2.
- an immunogenic composition comprises a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof; and an antigen from Neissiria gonorrhoeae. ).
- an immunogenic composition comprises a chlamydia polypeptide antigen selected from Table 1 , Table 2, Table 3, or combinations thereof; and an antigen from Candida albicans.
- an immunogenic composition comprises a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof; and an antigen from one or more of a papillomavirus, a herpesvirus (e.g., herpes simplex virus-2), Neissiria gonorrhoeae, and Candida albicans
- a herpesvirus e.g., herpes simplex virus-2
- Neissiria gonorrhoeae e.g., Neissiria gonorrhoeae
- Candida albicans e.g., Candida albicans
- the invention provides isolated nucleic acids comprising a nucleotide sequence encoding a chlamydia antigen described herein. In some embodiments, the invention provides isolated nucleic acids comprising a nucleotide sequence encoding a chlamydia antigen selected from Table 1, Table 2, Table 3, or combinations thereof. In some embodiments, a nucleic acid further comprises a nucleotide sequence encoding a
- heterologous peptide fused to the chlamydia antigen.
- compositions including nucleic acids encoding a chlamydia antigen as described herein.
- a composition includes an isolated nucleic acid comprising a nucleotide sequence encoding a chlamydia antigen selected from Table 1, Table 2, Table 3, or combinations thereof, and further comprises a pharmaceutically acceptable excipient.
- a composition further comprises an adjuvant.
- the invention provides methods for eliciting an immune response against chlamydia in a mammal based on nucleic acids described herein.
- the invention provides methods for eliciting an immune response against chlamydia in a mammal by administering to the mammal a composition comprising a nucleic acid, wherein the nucleic acid comprises a nucleotide sequence encoding a chlamydia antigen selected from Table 1, Table 2, Table 3, or combinations thereof.
- the invention provides methods for characterizing and/or detecting an immune response to a chlamydia antigen in a subject (e.g., a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof).
- a chlamydia antigen in a subject e.g., a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof.
- an immune response in a naive subject is characterized.
- an immune response in a subject infected, or suspected of having been infected, with chlamydia is characterized.
- an immune response in a subject administered an immunogenic composition comprising a chlamydia antigen e.g., an immunogenic composition described herein
- an antibody response is characterized.
- a B cell response is characterized.
- a T cell response is characterized.
- IFN- ⁇ secretion by antigen- specific T cells is characterized.
- a Thl T cell response is characterized.
- a Thl7 T cell response is characterized.
- a cytotoxic T cell response is characterized.
- both a T cell and a B cell response are characterized.
- an innate immune response is characterized.
- the invention further provides methods of preparing compositions including chlamydia antigens, and antibodies that specifically bind to chlamydia antigens.
- compositions and methods described herein can be used for the prophylaxis and/or treatment of any chlamydial disease, disorder, and/or condition, e.g., any of urethritis, cervicitis, pharyngitis, proctitis, epididymitis, prostatitis, pelvic inflammatory disease, and trachoma, due to a chlamydia infection.
- an immunogenic composition described herein reduces risk of infection by, and/or treats, alleviates, ameliorates, relieves, delays onset of, inhibits progression of, reduces severity of, and/or reduces incidence of one or more symptoms or features of a chlamydial disease, disorder, and/or condition.
- the prophylaxis and/or treatment of chlamydia infection comprises
- a "therapeutically effective amount" of an inventive immunogenic composition is that amount effective for treating, alleviating, ameliorating, relieving, delaying onset of, inhibiting progression of, reducing severity of, and/or reducing incidence of one or more symptoms or features of chlamydia infection.
- inventive prophylactic, prognostic and/or therapeutic protocols involve administering a therapeutically effective amount of one or more
- immunogenic compositions comprising a novel chlamydia antigen to a subject such that an immune response is stimulated in one or both of T cells and B cells.
- the present invention provides novel immunogenic compositions comprising a therapeutically effective amount of one or more chlamydia antigens (e.g., one or more of a polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof) and one or more pharmaceutically acceptable excipients.
- the present invention provides for pharmaceutical compositions comprising an immunogenic
- composition as described herein.
- a method of administering a pharmaceutical composition comprising inventive compositions to a subject e.g. human, e.g., a child, adolescent, or young adult in need thereof is provided.
- a therapeutically effective amount of an immunogenic composition is delivered to a patient and/or animal prior to, simultaneously with, and/or after diagnosis with a chlamydial disease, disorder, and/or condition.
- a therapeutic amount of an inventive immunogenic composition is delivered to a patient and/or animal prior to, simultaneously with, and/or after onset of symptoms of a chlamydial disease, disorder, and/or condition.
- immunogenic compositions of the present invention are administered by any of a variety of routes, including oral, intramuscular, subcutaneous, transdermal, interdermal, rectal, intravaginal, mucosal, nasal, buccal, enteral, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
- routes including oral, intramuscular, subcutaneous, transdermal, interdermal, rectal, intravaginal, mucosal, nasal, buccal, enteral, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
- immunogenic compositions of the present invention are administered by a variety of routes, including intravenous, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), transdermal, or by intratracheal instillation.
- routes including intravenous, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), transdermal, or by intratracheal instillation.
- an immunogenic composition may be administered in combination with one or more additional therapeutic agents which treat the symptoms of chlamydia infection (e.g., with an antibiotic such as an erythromycin or a tetracycline).
- additional therapeutic agents which treat the symptoms of chlamydia infection (e.g., with an antibiotic such as an erythromycin or a tetracycline).
- kits comprising one or more of the immunogenic compositions of the invention.
- the invention provides a kit comprising an immunogenic composition comprising a chlamydia antigen, or a nucleic acid encoding the antigen, wherein the antigen is selected from Table 1, Table 2, Table 3, or combinations thereof; and instructions for use.
- a kit may comprise multiple different chlamydia antigens.
- a kit may comprise any of a number of additional components or reagents in any combination.
- a kit may include, for example, (i) a chlamydia polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof; (ii) an adjuvant; and (iii) instructions for administering a composition including the chlamydia antigen and the adjuvant to a subject in need thereof.
- FIG. 1, 2, and 3 depict exemplary graphs illustrating the frequency with which identified antigens were recognized by human donor CD4 + and CD8 + T cells, respectively.
- Human donors were women with documented Chlamydia trachomatis exposure or a clinical history of genital infection. Donors were classified as "protected” if they were repeatedly exposed to the bacteria but not infected, or if they became infected but cleared their infection without medical intervention. Donors were classified as "unprotected” if they were persistently infected or if their infections progressed to more severe complications such as pelvic inflammatory disease.
- a donor was classified as a "responder” if the fold ratio of the response value over negative control was greater than 1.63 (CD4 + ) or 1.66 (CD8 + ). Percent responders >10 indicated a higher number of responders than due to chance alone.
- FIG. 1 depicts an exemplary result for protected and unprotected donors.
- FIG. 2 depicts another exemplary result for protected and unprotected donors.
- Four C. trachomatis proteins induced CD4 + or CD8 + T cell responses (two clones each, respectively) with statistically greater frequency in protected compared to unprotected donors, with a p-value of 0.05.
- FIG. 3 depicts an exemplary result illustrating CD4 + , CD8 + , and combined T cell responses for all donors (protected and unprotected). Antigens represented at the highest overall frequency, whether or not represented at statistically higher frequency in protected donors, are also attractive candidates for vaccine, diagnostic and prognostic applications.
- FIG. 4 depicts an exemplary result illustrating the frequency with which chlamydia antigens were bound by IgG present in donor sera, i.e. have elicited a donor B cell response.
- the left side of the panel displays chlamydia antigens detected by IgG with overall highest frequency across all donors (protected and unprotected).
- the right side of the panel displays chlamydia antigens detected by IgG with statistically greater frequency in protected donors as compared to unprotected donors.
- FIG. 5 depicts an exemplary result illustrating IFN- ⁇ levels induced ex vivo in
- FIG. 5A depicts an exemplary result illustrating antigens that were originally identified through T cell responses.
- FIG. 5B depicts an exemplary result illustrating antigens that were originally identified through B cell responses, demonstrating that these antigens can in some cases also elicit robust T cell responses.
- FIG. 6 depicts an exemplary result illustrating IgG antibody titers against each chlamydia antigen, following immunization with the same antigen.
- Exemplary results shown in the left side of the panel illustrate that antigens originally identified through T cell responses (e.g. FIG. 1, 2 and 3) can in some cases also elicit robust B cell responses.
- FIG. 7 depicts an exemplary result illustrating reduction of ectocervical chlamydia burden in mice immunized with identified chlamydia protein antigens and subsequently intravaginally infected with Chlamydia trachomatis.
- FIG. 7A depicts an exemplary result for representative chlamydia protein antigens CT062, CT043, and for the combination CT062 + CT043.
- FIG. 7B depicts an exemplary result for representative chlamydia protein antigen combination CT638 + CT476.
- FIG. 8 depicts an exemplary result illustrating reduction of upper reproductive tract chlamydia burden in mice immunized with the identified chlamydia protein antigens and subsequently intravaginally infected with Chlamydia trachomatis.
- FIG. 8A depicts an exemplary result for representative chlamydia protein antigens CT062, CT043, and for the combination CT062 + CT043.
- UVEB indicates responses from mice immunized with the positive control, UV-inactivated whole Chlamydia trachomatis elementary bodies.
- FIG. 8B depicts an exemplary result for representative chlamydia protein antigens CT067, CT0788tm, and CT328.
- FIG. 9 depicts an exemplary result illustrating induction of IFN- ⁇ in CD4 +
- Adjuvant As used herein, the term “adjuvant” refers to an agent that alters
- an adjuvant is used to enhance an immune response to a peptide antigen administered to a subject. In some embodiments, an adjuvant is used to enhance an immune response to an antigen encoded by a nucleic acid administered to a subject.
- Antibody refers to any immunoglobulin, whether natural or wholly or partially synthetically produced. All derivatives thereof which maintain specific binding ability are also included in the term. The term also covers any protein having a binding domain which is homologous or largely homologous to an immunoglobulin binding domain. Such proteins may be derived from natural sources, or partly or wholly synthetically produced. An antibody may be monoclonal or polyclonal. An antibody may be a member of any immunoglobulin class, including any of the human classes: IgG, IgM, IgA, IgD, and IgE. As used herein, the terms "antibody fragment" or
- characteristic portion of an antibody are used interchangeably and refer to any derivative of an antibody which is less than full-length.
- an antibody fragment retains at least a significant portion of the full-length antibody' s specific binding ability.
- antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, scFv, Fv, dsFv diabody, and Fd fragments.
- An antibody fragment may be produced by any means. For example, an antibody fragment may be enzymatically or chemically produced by
- an antibody fragmentation of an intact antibody and/or it may be recombinantly produced from a gene encoding the partial antibody sequence.
- an antibody fragment may be wholly or partially synthetically produced.
- An antibody fragment may optionally comprise a single chain antibody fragment.
- an antibody fragment may comprise multiple chains which are linked together, for example, by disulfide linkages.
- An antibody fragment may optionally comprise a multimolecular complex.
- a functional antibody fragment will typically comprise at least about 50 amino acids and more typically will comprise at least about 200 amino acids.
- Antigen specific immunological responses are mediated by lymphocytes (e.g., T cells, B cells) that express antigen receptors (e.g., T cell receptors, B cell receptors).
- an antigen is a T cell antigen, and elicits a cellular immune response.
- an antigen is a B cell antigen, and elicits a humoral (i.e., antibody) response.
- an antigen is both a T cell antigen and a B cell antigen.
- antigen encompasses both a full-length polypeptide as well as a portion of the polypeptide, that represent immunogenic fragments (i.e., fragments that elicit an antigen specific T cell response, B cell response, or both) of such complete polypeptides.
- antigen is a peptide epitope found within a polypeptide sequence (e.g., a peptide epitope bound by a Major Histocompatibility Complex (MHC) molecule (e.g., MHC class I, or MHC class II).
- MHC Major Histocompatibility Complex
- peptides 5-15 amino acids in length, and longer polypeptides, e.g., having 60, 70, 75, 80, 85, 90, 100, 150, 200 250, or more amino acids, can be "antigens".
- the present invention provides a CT062 polypeptide antigen.
- a CT062 polypeptide antigen includes a full-length CT062
- a CT062 polypeptide antigen includes a portion of a CT062 polypeptide (e.g., a portion of the CT062 polypeptide of SEQ ID NO: l, which portion includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 contiguous amino acids of SEQ ID NO: l).
- a CT062 polypeptide antigen contains one or more amino acid alterations (e.g., deletion, substitution, and/or insertion) from a naturally- occurring wild-type CT062 polypeptide sequence.
- a CT062 polypeptide antigen may contain an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: l or a portion thereof (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l).
- a CT062 polypeptide antigen may contain a portion (e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids) of a sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: l.
- CT062 polypeptide antigen is used as an example. This concept is applicable to other polypeptide antigen described herein including, but not limited to, CT572, CT043, CT570, CT177, CT725, CT067, CT476, p6, CT310, and CT638 polypeptide antigens.
- Chlamydia antigen refers to an antigen that elicits an antigen specific immune response against any organism of the
- Chlamydia genus such as a Chlamydia trachomatis organism, a Chlamydia psittaci organism, or a Chlamydia pneumoniae organism, a Chlamydia suis organism, a Chlamydia muridarum organism, etc.
- a chlamydia antigen elicits an antigen specific immune response against chlamydia organisms of multiple species (e.g., two or three of Chlamydia trachomatis, Chlamydia psittaci, and Chlamydia pneumoniae).
- a chlamydia antigen elicits an antigen specific immune response against chlamydia organisms of multiple serovars (e.g., one or more of serovars A, B, Ba, C, D, E, F, G, H, I, J, K, LI, L2, L3 of C. trachomatis).
- Chlamydia antigens include full-length polypeptides encoded by chlamydia genes, as well as immunogenic portions of the polypeptides.
- Immunogenic composition refers to a composition that includes a molecule that induces an immune response in a subject.
- an immunogenic composition includes a polypeptide or peptide antigen.
- an immunogenic composition includes a nucleic acid encoding a polypeptide or peptide antigen.
- An immunogenic composition can include molecules that induce an immune response against multiple antigens.
- in vitro refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within an organism (e.g., animal, plant, and/or microbe).
- in vivo refers to events that occur within ganism (e.g., animal, plant, and/or microbe).
- Isolated means that the isolated entity has been separated from at least one component with which it was previously associated. When most other components have been removed, the isolated entity is "purified.” Isolation and/or purification and/or concentration may be performed using any techniques known in the art including, for example, chromatography, fractionation, precipitation, or other separation.
- nucleic acid As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and/or substance that is or can be incorporated into an
- a nucleic acid is a compound and/or substance that is or can be incorporated into an oligonucleotide chain via a phosphodiester linkage.
- oligonucleotide and “polynucleotide” can be used interchangeably.
- nucleic acid encompasses RNA as well as single and/or double- stranded DNA and/or cDNA.
- nucleic acid “DNA,” “RNA,” and/or similar terms include nucleic acid analogs, i.e. analogs having other than a phosphodiester backbone.
- nucleotide sequence encoding an amino acid sequence includes all nucleotide sequences that are degenerate versions of each other and/or encode the same amino acid sequence.
- Nucleic acids can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, nucleic acids can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, etc.
- a nucleic acid sequence is presented in the 5' to 3' direction unless otherwise indicated.
- Polypeptide generally has its art- recognized meaning of a polymer of at least three amino acids. However, the term is also used to refer to specific classes of antigen polypeptides, such as, for example, CT062 polypeptides, CT572 polypeptides, CT043 polypeptides, CT570 polypeptides, CT177 polypeptides, and CT725 polypeptides. For each such class, the present specification provides several examples of known sequences of such polypeptides.
- polypeptide as used herein to refer to "polypeptide antigen” is intended to be sufficiently general as to encompass not only polypeptides having a sequence recited herein, but also to encompass polypeptides having a variation of the sequence that elicits an antigen- specific response to the polypeptide.
- a "CT062 polypeptide” includes the CT062 polypeptide shown in SEQ ID NO: l, as well as polypeptides that have variations of a SEQ ID NO: 1 sequence and that maintain the ability to elicit an antigen-specific response to a polypeptide of SEQ ID NO: 1.
- BLAST is used, with the parameters described herein, to determine percent sequence identity for the nucleic acids and proteins of the present disclosure.
- Software for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (available at the following internet address: ncbi.nlm.nih.gov).
- HSPs high scoring sequence pairs
- T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always>0) and N (penalty score for mismatching residues; always ⁇ 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score.
- Extension of the word hits in each direction are halted when: the cumulative alisnment score falls off bv the auantitv X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative- scoring residue alignments; or the end of either sequence is reached.
- the BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment.
- the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA, 90:5873- 5787, 1993).
- One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance.
- P(N) the smallest sum probability
- a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.
- Subject refers to any organism to which a composition of this invention may be administered, e.g., for
- Typical subjects include mammals such as mice, rats, rabbits, non-human primates, and humans.
- a disease, disorder, and/or condition has not been diagnosed with and/or may not exhibit symptoms of the disease, disorder, and/or condition.
- a disease, disorder, and/or condition is associated with a chlamydia infection (e.g., a C. trachomatis infection, a C. pneumoniae infection, or a C. psittaci infection).
- a chlamydia infection e.g., a C. trachomatis infection, a C. pneumoniae infection, or a C. psittaci infection.
- an individual who is susceptible to a chlamydia infection may be exposed to a chlamydia microbe (e.g., by ingestion, inhalation, physical contact, etc.).
- an individual who is susceptible to a chlamydia infection may be exposed to an individual who is infected with the microbe.
- an individual who is susceptible to a chlamydia infection is one who is in a location where the microbe is prevalent (e.g., one who is traveling to a location where the microbe is prevalent).
- an individual who is susceptible to a chlamydia infection is susceptible due to young age (e.g., a child, adolescent, or young adult).
- an individual who is susceptible to a disease, disorder, and/or condition will develop the disease, disorder, and/or condition.
- an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition.
- therapeutically effective amount means an amount of a therapeutic, prophylactic, and/or diagnostic agent (e.g., inventive immunogenic composition) that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and/or condition, to treat, alleviate, ameliorate, relieve, alleviate symptoms of, prevent, delay onset of, inhibit progression of, reduce severity of, and/or reduce incidence of the disease, disorder, and/or condition.
- a therapeutic, prophylactic, and/or diagnostic agent e.g., inventive immunogenic composition
- Therapeutic agent refers to any agent that, when administered to a subject, has a therapeutic, prophylactic, and/or diagnostic effect and/or elicits a desired biological and/or pharmacological effect.
- Treating refers to partially or completely alleviating, ameliorating, relieving, delaying onset of, inhibiting progression of, reducing severity of, and/or reducing incidence of one or more symptoms or features of a particular disease, disorder, and/or condition.
- treating a microbial infection may refer to inhibiting survival, growth, and/or spread of the microbe.
- Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition and/or to a subject who exhibits only early signs of a disease, disorder, and/or condition for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
- treatment comprises delivery of an immunogenic composition (e.g., a vaccine) to a subject.
- Vaccine refers to an entity comprising at least one immunogenic component (e.g., an immunogenic component which includes a peptide or protein, and/or an immunogenic component which includes a nucleic acid).
- a vaccine includes at least two immunogenic components.
- a vaccine is capable of stimulating an immune response of both T cells and B cells.
- any assay available in the art may be used to determine whether T cells and/or B cells have been stimulated.
- T cell stimulation may be assayed by monitoring antigen-induced production of cytokines, antigen-induced
- B cell stimulation may be assayed by monitoring antibody titers, antibody affinities, antibody performance in neutralization assays, class-switch recombination, affinity maturation of antigen- specific antibodies, development of memory B cells, development of long-lived plasma cells that can produce large amounts of high-affinity antibodies for extended periods of time, germinal center reactions, and/or antibody performance in neutralization assays.
- a vaccine further includes at least one adjuvant that can help stimulate an immune response in T cells and/or B cells.
- Wild-type As used herein, the term “wild-type” refers to the typical or the most common form existing in nature.
- Infection by Chlamydia trachomatis causes inflammation and damage to mucosal tissues, leading to pathologies such as urethritis, cervicitis, pharyngitis, proctitis, epididymitis, prostatitis, and trachoma, and infertility secondary to these pathologies.
- Chlamydia bacteria which primarily infect epithelial cells, alternate between two
- EB forms of chlamydia are infectious and invade host cells. After forming an inclusion within host cells, EB forms differentiate into RB forms which replicate for a period of time and differentiate back to EB forms.
- C. trachomatis species are categorized into serovars based on reactivity of patient sera to the major outer membrane protein (MOMP).
- MOMP major outer membrane protein
- Serovars A, B, Ba, and C are associated with infection of conjunctival epithelium.
- Serovars D-K are associated with urogenital tract infections.
- Serovars L1-L3 are associated with urogenital tract infection and a systemic condition, lymphogranuloma venereum.
- CD4 + T cell responses of the Thl subtype have been shown to be important for clearance of chlamydia infections in an animal model (Morrison et al., Infect. Immun. 70:2741-2751, 2002). B cell responses are thought to contribute to protective immunity in humans and non-human primates (Brunham et al., Infect. Immun. 39: 1491-1494, 1983; Taylor et al., Invest. Ophthalmol. Vis. Sci 29: 1847-1853, 1988). CD8 + T cells have lytic functions that are important for the control of intracellular pathogens.
- Chlamydia-specific CD8 + T cells have been isolated from infected humans, indicating a role for these cells in responding to chlamydia infections (Gervassi et al., J. Immunol. 171: 4278- 4286, 2003).
- the present invention provides chlamydia antigens, including, but not limited to, CT062 polypeptide antigens, CT572 polypeptide antigens, CT043 polypeptide antigens, CT570 polypeptide antigens, CT177 polypeptide antigens, CT725 polypeptide antigens, CT067 polypeptide antigens, CT476 polypeptide antigens, p6 polypeptide antigens, CT310 polypeptide antigens, and CT638 polypeptide antigens that are recognized by immune cells (e.g., T cells and/or B cells) of infected mammals. As described in the Examples herein, these antigens were discovered as targets of T cell- or B cell-mediated immunity in vivo.
- immune cells e.g., T cells and/or B cells
- these antigens provide novel compositions for eliciting immune responses with the aim of eliciting beneficial immune responses, e.g., to protect against chlamydia infections and associated pathologies. These antigens also provide novel targets for characterizing chlamydia infections and immune responses to chlamydia infections.
- CT062 polypeptides are cytoplasmic tyrosyl-tRNA synthetases in chlamydia organisms. Exemplary amino acid and nucleotide sequences from a full-length CT062 polypeptide of C. trachomatis are shown below as SEQ IDs NO: 1 and 2. In some
- a CT062 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of a CT062 polypeptide sequence, e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l or of a sequence at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO: l.
- a CT062 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, or 400 consecutive amino acids of the sequence shown in SEQ ID NO: l.
- a CT062 polypeptide antigen is a full-length CT062 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO: l).
- a CT062 polypeptide antigen lacks one or more trans-membrane domains (e.g., a CT062 polypeptide antigen lacks amino acids 55-74 of SEQ ID NO: l).
- CT572 polypeptides are known as general secretion pathway proteins D.
- a CT572 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of a CT572 polypeptide sequence, e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3 or of a sequence at least 60% (e.g., at least
- a CT572 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, or 750 consecutive amino acids of the sequence shown in SEQ ID NO:3.
- a CT572 polypeptide antigen is a full-length CT572 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:3).
- a CT572 polypeptide antigen lacks one or more trans-membrane domains and/or a signal sequence (e.g., a CT572 polypeptide antigen lacks amino acids 1-24 of SEQ ID NO:3).
- a CT043 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of a CT043 polypeptide sequence, e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5 or of a sequence at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO:5.
- a CT043 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, or 160 consecutive amino acids of the sequence shown in SEQ ID NO:5.
- a CT043 polypeptide antigen is a full-length CT043 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:5).
- a CT043 polypeptide antigen lacks one or more trans-membrane domains (e.g., a CT043 polypeptide antigen lacks amino acids 75-93 of SEQ ID NO:5).
- CT570 polypeptides are known as general secretion pathway proteins F.
- a CT570 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of a CT570 polypeptide sequence, e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7 or of a sequence at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO:7.
- a CT570 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 350 consecutive amino acids of the sequence shown in SEQ ID NO:7.
- a CT570 polypeptide antigen is a full-length CT570 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:7).
- a CT570 polypeptide antigen lacks one or more transmembrane domains (e.g., a CT570 polypeptide antigen lacks amino acids 164-182 and/or 211-230 and/or 363-382 of SEQ ID NO:7).
- CT177 polypeptides are disulfide bond chaperone proteins. Exemplary amino acid and nucleotide sequences from a full-length CT177 polypeptide of C. trachomatis are shown below as SEQ IDs NO:9 and 10.
- a CT177 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of a CT177 polypeptide sequence, e.g., at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9 or of a sequence at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO:9.
- a CT177 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 100, 150, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:9.
- a CT177 polypeptide antigen is a full-length CT177 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:9).
- a CT177 polypeptide antigen lacks one or more trans-membrane domains and/or a signal sequence (e.g., a CT177 polypeptide antigen lacks amino acids 1-30 of SEQ ID NO:9).
- CT725 polypeptides are biotin synthetases. Exemplary amino acid and nucleotide sequences from a full-length CT725 polypeptide of C. trachomatis are shown below as SEQ IDs NO: 11 and 12.
- a CT725 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of a CT725 polypeptide sequence, e.g.
- a CT725 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 120, 130, 140, 150, 160, 170, or 180 consecutive amino acids of the sequence shown in SEQ ID NO: 11.
- a CT725 polypeptide antigen is a full-length CT725 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO: 11).
- a CT726 polypeptide antigen lacks one or more trans-membrane domains (e.g., a CT726 polypeptide antigen lacks amino acids 51-75 and/or 116-136 of SEQ ID NO: 11).
- CT067 polypeptides are ABC transporter proteins. Exemplary amino acid and nucleotide sequences from a full-length CT067 polypeptide of C. trachomatis are shown below as SEQ IDs NO:23 and 24.
- a CT067 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of a CT067 polypeptide sequence, e.g.
- a CT067 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 325 consecutive amino acids of the sequence shown in SEQ ID NO:23.
- a CT067 polypeptide antigen is a full-length CT067 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:23).
- a CT067 polypeptide antigen lacks one or more trans-membrane domains and/or a signal sequence (e.g., a CT067 polypeptide antigen lacks amino acids 1-33 and/or amino acids 11-31 of SEQ ID NO:23).
- CT476 polypeptides are of unknown function. Exemplary amino acid and nucleotide sequences from a full-length CT476 polypeptide of C. trachomatis are shown below as SEQ IDs NO:63 and 64.
- a CT476 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of a CT476 polypeptide sequence, e.g.
- a CT476 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, 250, 300, or 320 consecutive amino acids of the sequence shown in SEQ ID NO:63.
- a CT476 polypeptide antigen is a full-length CT476 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:63).
- a CT476 polypeptide antigen lacks one or more trans-membrane domains and/or a signal sequence (e.g., a CT476 polypeptide antigen lacks amino acids 1-18 and/or amino acids 1-20 of SEQ ID NO:63).
- Chlamydia p6 polypeptides are plasmid virulence factors PGP4-D.
- a p6 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of a p6 polypeptide sequence, e.g.
- a p6 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, or 100 consecutive amino acids of the sequence shown in SEQ ID NO:65.
- a p6 polypeptide antigen is a full-length p6 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:65).
- a p6 polypeptide antigen lacks one or more trans-membrane domains (e.g., a p6 polypeptide antigen lacks amino acids 52-68 of SEQ ID NO:65).
- CT310 polypeptides are putative ATP synthase subunits. Exemplary amino acid and nucleotide sequences from a full-length CT310 polypeptide of C. trachomatis are shown below as SEQ IDs NO:67 and 68.
- a CT310 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 160, 170, 180, 190, or 200 consecutive amino acids of a CT310 polypeptide sequence, e.g.
- a CT310 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 160, 170, 180, 190, or 200 consecutive amino acids of the sequence shown in SEQ ID NO:67.
- a CT310 polypeptide antigen is a full-length CT310 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:67).
- a CT310 polypeptide antigen lacks one or more transmembrane domains (e.g., a CT310 polypeptide antigen lacks amino acids 117-136 of SEQ ID NO:67).
- CT638 polypeptides are of unknown function. Exemplary amino acid and nucleotide sequences from a full-length CT638 polypeptide of C. trachomatis are shown below as SEQ IDs NO:69 and 70.
- a CT638 polypeptide antigen includes at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 250 consecutive amino acids of a CT638
- polypeptide sequence e.g. at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69 or of a sequence at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to SEQ ID NO:69.
- a CT638 polypeptide antigen comprises an amino acid sequence that is at least 60% (e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 98%) identical to at least 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 45, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 250 consecutive amino acids of the sequence shown in SEQ ID NO:69.
- a CT638 polypeptide antigen is a full-length CT310 polypeptide (e.g., the antigen comprises the amino acid sequence of SEQ ID NO:69).
- a CT638 polypeptide antigen lacks one or more trans-membrane domains and/or a signal sequence (e.g., a CT638 polypeptide antigen lacks amino acids 1-33 and/or amino acids 13- 31 of SEQ ID NO:69).
- CT757, CT564, CT703, P1-ORF7, CT067, CT037, CT252, CT064, CT137, CT204, CT634, CT635, CT366, CT140, CT142, CT242, CT843, CT328, CT188, CT578, CT724, CT722, CT732, and CT788 polypeptide antigens are shown below as SEQ IDs NO: 13-62.
- Exemplary amino acid and nucleotide sequences from full-length CT172, CT443, CT525, CT606, CT648, and CT870 polypeptide antigen are shown below as SEQ IDs NO:71-82.
- Polypeptide antigens of Table 1 can be provided in any combination with each other and/or with other chlamydia antigens.
- a combination of chlamydia polypeptide antigens includes two polypeptide antigens selected from Table 1.
- a combination includes three polypeptide antigens selected from Table 1.
- a combination includes four polypeptide antigens selected from Table 1.
- a combination includes five polypeptide antigens selected from Table 1.
- a combination includes six polypeptide antigens selected from Table 1.
- a combination includes seven polypeptide antigens selected from Table 1.
- a combination includes eight polypeptide antigens selected from Table 1.
- antigens which can be provided in combination with one or more polypeptide antigens selected from Table 1 include one or more polypeptide antigens selected from Table 2 , and/or one or more polypeptide antigens selected from Table 3.
- a combination of antigens includes one, two, three, four, five, six, seven, or eight polypeptide antigens selected from Table 1, and one, two, three, four, five, or six polypeptide antigens selected from Table 2.
- a combination of antigens includes one, two, three, four, five, six, seven, or eight polypeptide antigens selected from Table 1, and one, two, three, four, five, or six polypeptide antigens selected from Table 3.
- a combination of antigens includes one, two, three, four, five, six, seven, or eight polypeptide antigens selected from Table 1; one, two, three, four, five, or six polypeptide antigens selected from Table 2; and one, two, three, four, five, or six polypeptide antigens selected from Table 3.
- a combination of antigens includes one, two, three, four, five, or six polypeptide antigens selected from Table 2, and one, two, three, four, five, or six polypeptide antigens selected from Table 3.
- Antigens CT062, CT843, CT242, CT732, CT788, and specific epitopes of these antigens are described in
- PCT/US2007/004675 (published as WO 2007/098255), PCT/US2008/009282 (published as WO 2009/020553), PCT/US2008/013298 (published as WO 2009/073179), and
- Additional chlamydia polypeptide antigens that can be provided in combination with a novel antigen described herein include a polymorphic membrane protein D (PmpD or CT812; see GenBank NP_220332.1 GI: 15605546), a major outer membrane protein (MOMP or ompA or CT681; see GenBank NP_220200.1
- PmpD or CT812 polymorphic membrane protein D
- MOMP or ompA or CT681 see GenBank NP_220200.1
- compositions that include a chlamydia antigen described herein and an antigen from a different infectious agent.
- a composition includes a chlamydia antigen and an antigen from a different infectious agent that causes a sexually transmitted disease.
- compositions that include a chlamydia antigen e.g., a polypeptide antigen selected from Table 1, Table 2, Table 3, or a combination thereof
- a papillomavirus antigen e.g., a human papillomavirus antigen
- compositions that include a chlamydia antigen e.g., a polypeptide antigen selected from Table 1, Table 2, Table 3, or a combination thereof
- a herpesvirus antigen e.g., a human herpes simplex virus-2 antigen
- compositions that include a chlamydia antigen (e.g., a polypeptide antigen selected from Table 1, Table 2, Table 3, or a combination thereof) and a Neissiria gonorrhoea antigen are provided.
- compositions that include a chlamydia antigen e.g., a polypeptide antigen selected from Table 1. Table 2. Table 3.
- compositions that include a chlamydia antigen (e.g., a polypeptide antigen selected from Table 1, Table 2, Table 3, or a combination thereof) and an antigen from one or more of a papillomavirus, a herpesvirus (e.g., HSV-2), Neissiria gonorrhoeae, and Candida albicans are provided.
- a chlamydia antigen e.g., a polypeptide antigen selected from Table 1, Table 2, Table 3, or a combination thereof
- an antigen from one or more of a papillomavirus e.g., HSV-2
- Neissiria gonorrhoeae e.g., a gonorrhoeae
- Candida albicans e.g., Candida albicans.
- immunogenic compositions can be employed to induce immune responses.
- a common route of administration in humans is by intramuscular (i.m.) injection, but immunogenic compositions may also be applied orally, intranasally, subcutaneously, by inhalation, intravenously, or by other routes of
- lymphocytes in regional lymph nodes are lymphocytes in regional lymph nodes.
- a chlamydia antigen composition includes purified components (e.g., purified antigens).
- chlamydia antigens are fused to other molecules, such as proteins that can confer adjuvant activity, or moieties that facilitate isolation and purification (e.g., an epitope tag).
- a chlamydia antigen composition includes an adjuvant.
- the adjuvant includes mineral-containing adjuvant.
- Mineral- containing ajduvants can be formulated as gels, in crystalline form, in amorphous form, as particles, etc.
- Mineral-containing adjuvants include, for example, aluminum salts and/or calcium salts (e.g., aluminum hydroxide, aluminum phosphate, aluminum sulfate, calcium phosphate, etc.).
- a chlamydia antigen composition includes aluminum hydroxide.
- AlhydrogelTM is an example of an aluminum hydroxide gel adjuvant.
- an adjuvant includes an immunomodulatory oligonucleotide.
- an immunomodulatory oligonucleotide sequence includes CpG (unmethylated cytosine-guanosine) motifs.
- Oligonucleotides having CpG motifs can include nucleotide analogs and/or non-naturally occuring intemucleoside linkages (e.g., phosphorothioate linkages).
- CpG motifs see Kandimalla, et al., Nuc. Acids Res. 31(9): 2393-2400, 2003; WO02/26757;
- an adjuvant comprises IC31TM (Intercell AG).
- IC31TM is a synthetic adjuvant that includes an antimicrobial peptide, KLK, and an
- TLR9 Toll-like Receptor 9
- an adjuvant includes a toxin.
- a toxin is a bacterial ADP-ribosylating toxin, e.g., cholera toxin, E. coli heat labile toxin, or pertussis toxin.
- the bacterial toxin is a detoxified form of an ADP- ribosylating toxin (see, e.g., Beignon, et al., Inf. Immun. 70(6):3012-3019, 2002; Pizza, et al., Vaccine 19:2534-2541, 2001; Pizza, et al., Int. J. Med. Microbiol. 290(4-5):455-461, 2000; Scharton-Kersten et al., Inf. Immun.68(9):5306-5313, 2000; Ryan et al., Inf. Immun.
- an adjuvant includes an endotoxin such as
- an adjuvant includes a muramyl dipeptide (e.g., N- acetyl-muramyl-L-threonyl-D-isoglutamine(thr-MDP), N-acetyl-normuramyl- 1 -alanyl-d- isoglutamine(nor-MDP), and N-acetylmuramyl-l-alanyl-d-isoglutaminyl-l-alanine-2-(l'-2'- dipalmitoyl- s - n- glycero- 3 -hydroxypho sphoryloxy) -ethylamine MTP-PE) .
- a muramyl dipeptide e.g., N- acetyl-muramyl-L-threonyl-D-isoglutamine(thr-MDP)
- an adjuvant includes an oil emulsion and/or emulsifier-based adjuvant.
- an oil emulsion adjuvant includes a Freund's Adjuvant (e.g., Complete Freund's adjuvant (CFA), or incomplete Freund's adjuvant (IFA)).
- an oil-emulsion adjuvant includes a squalene water emulsion, such as MF59 (Novartis; see, e.g., WO9014837), or a Synex adjuvant formulation (SAF)).
- an oil emulsion includes a dispersing agent, e.g., a mono- or di-C 1 2-C 2 4-fatty acid ester of sorbitan or mannide, e.g., sorbitan mono-stearate, sorbitan mon-oleate, or mannide mono-oleate.
- a dispersing agent e.g., a mono- or di-C 1 2-C 2 4-fatty acid ester of sorbitan or mannide, e.g., sorbitan mono-stearate, sorbitan mon-oleate, or mannide mono-oleate.
- examples of oil emulsions that include squalene and dispersing agents includes ArlacelTM, MontanideTM ISA-720, and MontanideTM ISA-703. Other oil emulsions are described, e.g., in WO 95/17210 and EP 0399842.
- an adjuvant includes a saponin.
- Saponins are steroid and/or triterpenoid glycosides derived from plants such as Quillaja saponaria, Saponaria officianalis, Smilax ornata, and Gypsophilla paniculata.
- Fractions of saponin-containing extracts that have been described and that can be used as adjuvants for chlamydia antigens include QuilTMA, QS21, QS7, QS17, QS18, QH-A, QH-B, QH-C, and QuilA (see, e.g., U.S. Pat. No. 5,057,540).
- QS21 is used as an adjuvant.
- an adjuvant includes an immune stimulating complex
- ISCOMs are particles that typically include a glycoside (e.g., a saponin) and a lipid.
- an ISCOM includes a saponin and a cholesterol.
- an ISCOM includes a saponin, a cholesterol, and a phospholipid (e.g., phosphatidylcholine and/or phosphatidylethanolamine).
- an ISCOM includes a nonionic block copolymer.
- ISCOMs can include additional adjuvants, e.g., additional adjuvant substances described herein (see, e.g., WO 05/002620).
- an ISCOM includes a substance that targets it to a mucosal membrane (see, e.g., WO97/030728).
- Other ISCOM compositions and preparation of the compositions suitable for combination with chlamydia antigens provided herein are described, e.g., in U.S. Pat. Pub. No. 20060121065, WO 00/07621, WO 04/004762, WO 02/26255, and WO 06/078213.
- an adjuvant comprises an AblSCO ® adjuvant (e.g., Matrix-MTM, Isconova).
- an adjuvant comprises AbISCO ® -100.
- an adjuvant comprises AbISCO ® -300.
- an adjuvant includes a nonionic block copolymer.
- Nonionic block copolymers typicaly include two chains of hydrophobic polyoxyethylenes of various lengths combined with a block of hydrophobic polyoxypropylene.
- a nonionic block copolymer is formulated in an oil-in-water emulsion (e.g., with oil and squalene).
- an adjuvant includes virus like particles (VLPs).
- VLPs are non replicating, non infectious particles that typically include one or more viral proteins, optionally formulated with an additional component such as a phospholipid.
- a VLP includes proteins from one or more of the following: an influenza virus (e.g., a hemaglutinin (HA) or neuraminidase (NA) polyptide), Hepatitis B virus (e.g., a core or capsid polypeptide), Hepatitis E virus, measles virus, Sindbis virus, Rotavirus, Foot-and- Mouth Disease virus, Retrovirus, Norwalk virus, human papilloma virus, HIV, RNA-phages, Q -phage (e.g., a coat protein), GA-phage, fr-phage, AP205 phage, a Ty (e.g., retrotransposon Ty protein pi). See, e.g., WO03/0244
- an adjuvant includes replicons. Replicons resemble
- VLPs in that they are noninfectious particles including viral proteins, and further include a nucleic acid encoding a polypeptide (e.g., an antigen).
- a replicon includes proteins from an alphavirus.
- Alphaviruses include, e.g., Eastern Equine Encephalitis Virus (EEE), Venezuelan Equine Encephalitis Virus (VEE), Everglades Virus, Mucambo Virus, Pixuna Virus, Western Equine Encephalitis Virus (WEE), Sindbis Virus, Semliki Forest Virus, Middleburg Virus, Chikungunya Virus, O'nyong-nyong Virus, Ross River Virus, Barmah Forest Virus, Getah Virus, Sagiyama Virus, Bebaru Virus, Mayaro Virus, Una Virus, Aura Virus, Whataroa Virus, Babanki Virus, Kyzylagach Virus, Highlands J Virus, Fort Morgan Virus, Ndumu Virus
- an adjuvant includes a replicon that includes a nucleic acid encoding one or more chlamydia antigens described herein.
- an adjuvant includes a replicon that encodes a cytokine (e.g., interleukin-12 (IL-12), IL-23, or granulocyte-macrophage colony- stimulating factor (GM-CSF)).
- IL-12 interleukin-12
- GM-CSF granulocyte-macrophage colony- stimulating factor
- a VLP or replicon adjuvant includes one or more chlamydia antigens (i.e., VLP or replicon particles include a chlamydia antigen as part of the particles).
- a VLP or replicon adjuvant is co-adminstered with a chlamydia antigen polypeptide.
- an adjuvant includes liposomes, which are are artificially-constructed spherical lipid vesicles (see, e.g., U.S. Pat. Nos. 4,053,585; 6,090,406; and 5,916,588).
- a lipid to be used in liposomes can be, but is not limited to, one or a plurality of the following: phosphatidylcholine, lipid A, cholesterol, dolichol, sphingosine, sphingomyelin, ceramide, glycosylceramide, cerebroside, sulfatide, phytosphingosine, phosphatidyl-ethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, cardiolipin, phosphatidic acid, and lyso-phosphatides.
- an adjuvant includes a liposome and a ligand for a Toll-like Receptor (TLR; see, e.g., WO/2005/013891, WO/2005/079511, WO/2005/079506, and WO/2005/013891).
- an adjuvant includes JVRS-100. JVRS-100 comprises cationic liposomes combined with non-coding oligonucleotides or plasmids.
- an adjuvant includes microparticles comprised of a polymer, e.g., a polymer of acrylic or methacrylic acid, polyphosphazenes, polycarbonates, polylactic acid, polyglycolic acid, copolymers of lactic acid or glycolic acid,
- a polymer e.g., a polymer of acrylic or methacrylic acid, polyphosphazenes, polycarbonates, polylactic acid, polyglycolic acid, copolymers of lactic acid or glycolic acid,
- an adjuvant includes microparticles comprised of a polymer selected from the group consisting of polyvinylpyrrolidone, polyvinylalcohol, polyhydroxyethylmethacrylate, polyacrylamide, polymethacrylamide, and polyethyleneglycol (see, e.g., U.S. Pat. No. 5,500,161).
- an adjuvant includes biodegradable microspheres (e.g., microspheres comprised of poly(D,L-lactic acid), poly(D,L-glycolic acid), poly(e- caprolactone), polye (cc-hydroxy actid), polyhydroxybutyric acid, a polyorthoester, a polyanhydride, etc.).
- biodegradable microspheres e.g., microspheres comprised of poly(D,L-lactic acid), poly(D,L-glycolic acid), poly(e- caprolactone), polye (cc-hydroxy actid), polyhydroxybutyric acid, a polyorthoester, a polyanhydride, etc.
- an adjuvant includes a cytokine.
- an adjuvant includes IL-12. In some embodiments, an adjuvant includes IL- 23. In some embodiments, an adjuvant includes GM-CSF.
- an adjuvant includes a lipopeptide. In some embodiments, an adjuvant includes a Pam-3-Cys lipopeptide. In some embodiments, an adjuvant including a lipopeptide activates Toll-like receptors (TLRs).
- TLRs Toll-like receptors
- chlamydia antigens described herein may be used with or without modification.
- a chlamydia antigen may be modified to elicit the desired immune response.
- a chlamydia antigen is conjugated to an appropriate immunogenic carrier such as tetatus toxin, pneumolysin, keyhole limpet hemocyanin, or the like.
- a chlamydia polypeptide antigen is post- translationally modified, e.g. by phosphorylation, myristoylation, acylation, glycosylation, glycation, and the like.
- a chlamydia polypeptide antigen is lipidated. Conjugation to the lipid moiety may be direct or indirect (e.g., via a linker). The lipid moiety may be synthetic or naturally produced. In some embodiments, a chlamydia polypeptide antigen is chemically conjugated to a lipid moiety. In some embodiments, a DNA construct encoding a chlamydia polypeptide antigen comprises a lipidation sequence. A lipidation sequence may be N-terminal or C-terminal to the polypeptide, and may be embedded in a signal or other sequence. An exemplary lipidation sequence is the signal sequence of the E. coli gene RlpB, shown as SEQ ID NO:83.
- a chlamydia polypeptide antigen is covalently bound to another molecule. This may, for example, increase the half-life, solubility, bioavailability, or immunogenicity of the antigen.
- Molecules that may be covalently bound to the antigen include a carbohydrate, biotin, poly(ethylene glycol) (PEG), polysialic acid, N-propionylated polysialic acid, nucleic acids, polysaccharides, and PLGA.
- the naturally produced form of a polypeptide is covalently bound to a moiety that stimulates the immune system.
- An example of such a moiety is a lipid moiety.
- lipid moieties are recognized by a Toll-like receptor (TLR) such as TLR2 or TLR4 and activate the innate immune system.
- TLR Toll-like receptor
- a composition includes a vector suitable for expression in vitro (whether in a cell or in a cell-free system), e.g., for producing a polypeptide composition.
- vector refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked and can include, for example, a plasmid, cosmid or viral vector.
- the vector can be capable of autonomous replication or it can integrate into a host DNA.
- Viral vectors include, e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses. Other types of viral vectors are known in the art.
- a vector can include a nucleic acid encoding a chlamydia antigen in a form suitable for expression of the nucleic acid in a host cell.
- a recombinant expression vector typically includes one or more regulatory sequences operatively linked to the nucleic acid sequence to be expressed. Regulatory sequences include promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Regulatory sequences include those which direct constitutive expression of a nucleotide sequence, as well as tissue-specific regulatory and/or inducible sequences.
- a sequence encoding a chlamydia antigen can include a sequence encoding a signal peptide (e.g., a heterologous signal peptide) such that the antigen is secreted from a host cell.
- a signal peptide e.g., a heterologous signal peptide
- the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, and the like.
- Recombinant expression vectors can be designed for expression and production of chlamydia antigens in prokaryotic or eukaryotic cells.
- antigens can be expressed in E. coli, insect cells (e.g., using baculovirus expression vectors), yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, Gene
- a recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.
- Fusion vectors add a number of amino acids to a protein encoded therein, e.g., to the amino terminus or carboxy terminus of the recombinant protein, e.g., to increase expression of recombinant protein; to increase the solubility of the recombinant protein; and/or to aid in the purification of the recombinant antigen by acting as a ligand in affinity purification.
- a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant antigen to enable separation of the recombinant antigen from the fusion moiety subsequent to purification of the fusion protein.
- enzymes, and their cognate recognition sequences include Factor Xa, thrombin and enterokinase.
- Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith, D.B. and Johnson, K.S.
- Chlamydia antigen expression vectors include yeast expression vectors, vectors for expression in insect cells (e.g., a baculovirus expression vector) and vectors suitable for expression in mammalian cells.
- An expression vector for use in mammalian cells can include viral regulatory elements.
- promoters are derived from polyoma, Adenovirus 2, cytomegalovirus and Simian Virus 40.
- a vector can include an inducible promoter, e.g., a promoter regulated by a steroid hormone, by a polypeptide hormone (e.g., by means of a signal transduction pathway), or by a heterologous polypeptide (e.g., the tetracycline- inducible systems, "Tet-On” and "Tet-Off '; see, e.g., Clontech Inc., CA, Gossen and Bujard, Proc. Natl. Acad. Sci. USA 89:5547, 1992, and Paillard, Human Gene Therapy 9:983, 1989).
- an inducible promoter e.g., a promoter regulated by a steroid hormone, by a polypeptide hormone (e.g., by means of
- a host cell can be any prokaryotic or eukaryotic cell.
- a chlamydia antigen can be expressed in bacterial cells (such as E. coli), insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells (African green monkey kidney cells CV-1 origin SV40 cells; Gluzman, Cell 23: 175-182, 1981).
- bacterial cells such as E. coli
- insect cells such as E. coli
- yeast or mammalian cells such as Chinese hamster ovary cells (CHO) or COS cells (African green monkey kidney cells CV-1 origin SV40 cells; Gluzman, Cell 23: 175-182, 1981).
- COS cells African green monkey kidney cells CV-1 origin SV40 cells; Gluzman, Cell 23: 175-182, 1981.
- Other suitable host cells are known to those skilled in the art.
- Vector DNA can be introduced into host cells via conventional transformation or transfection techniques.
- a host cell can be used to produce (i.e., express) a chlamydia antigen.
- the invention further provides methods for producing a chlamydia antigen using host cells.
- the method includes culturing a host cell (into which a recombinant expression vector encoding a chlamydia antigen has been introduced) in a suitable medium such that a chlamydia antigen is produced.
- the method further includes isolating a chlamydia antigen from the medium or the host cell. Purified chlamydia antigens can be used for administration to mammals to induce an immune response, and/or to generate antibodies specific for the antigens.
- the present invention also provides nucleic acid compositions that encode chlamydia antigens for administration to a subject in vivo, e.g., to elicit an immune response to the antigen.
- a nucleic acid composition for administration in vivo includes a naked DNA plasmid encoding a chlamydia antigen.
- Bacterial vectors, replicon vectors, live attenuated bacteria, and viral vectors for expression of heterologous genes also can be used.
- Live attenuated viral vectors e.g., recombinant vaccinia (e.g., modified vaccinia Ankara (MVA), IDT Germany), recombinant adenovirus, avian poxvirus (e.g., canarypox (e.g., ALVACTM, Aventis Pasteur) or fowlpox), poliovirus, and alphavirus virion vectors) have been successful in inducing cell-mediated immune response to antigens.
- Avian poxviruses are defective in mammalian hosts, but can express inserted heterologous genes under early promoters.
- Attenuated bacteria can also be used as a vehicle for DNA vaccine delivery.
- suitable bacteria include S. enterica, S. tymphimurium, Listeria, and BCG.
- S. enterica, S. tymphimurium, Listeria, and BCG suitable bacteria
- the use of mutant bacteria with weak cell walls can aid the exit of DNA plasmids from the bacterium.
- Nucleic acid compositions used for immunization can include an adjuvant
- an adjuvant such as a polymer, a saponin, muramyl dipeptide, liposomes,
- an adjuvant increases the uptake of nucleic acid into host cells and/or increases expression of the antigen from the nucleic acid within the cell, induce antigen presenting cells to infiltrate the region of tissue where the antigen is being expressed, or increase the antigen-specific response provided by
- lymphocytes are lymphocytes.
- This invention provides, inter alia, antibodies, or antigen-binding fragments thereof, to a novel chlamydia antigen described herein, e.g., a CT062 polypeptide antigen, a CT572 polypeptide antigen, a CT043 polypeptide antigen, a CT570 polypeptide antigen, a CT177 polypeptide antigen, a CT725 polypeptide antigen, a CT067 polypeptide antigen , a CT476 polypeptide antigen, a p6 polypeptide antigen, a CT310 polypeptide antigen, or a CT638 polypeptide antigen .
- a novel chlamydia antigen described herein e.g., a CT062 polypeptide antigen, a CT572 polypeptide antigen, a CT043 polypeptide antigen, a CT570 polypeptide antigen, a CT177 polypeptide antigen, a CT725 polypeptide antigen, a CT
- the antibodies can be of the various isotypes, including: IgG (e.g., IgGl, IgG2, IgG3, IgG4), IgM, IgAl, IgA2, IgD, or IgE.
- an antibody is an IgG isotype, e.g., IgGl.
- An antibody against a chlamydia antigen can be full- length (e.g., an IgGl or IgG4 antibody) or can include only an antigen-binding fragment (e.g., a Fab, F(ab)2, Fv or a single chain Fv fragment).
- These include monoclonal antibodies, recombinant antibodies, chimeric antibodies, human antibodies, and humanized antibodies, as well as antigen-binding fragments of the foregoing.
- Monoclonal antibodies can be produced by a variety of techniques, including conventional monoclonal antibody methodology, e.g., the standard somatic cell hybridization technique of Kohler and Milstein, Nature 256: 495, 1975. Polyclonal antibodies can be produced by immunization of animal or human subjects. See generally, Harlow, E. and Lane, D. Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1988. Antibodies against chlamydia antigens described herein can be used, e.g., for diagnostic assays, or for therapeutic applications.
- a subject's response to an immunogenic composition described herein is evaluated, e.g., to determine efficacy of the composition, and/or to compare responses elicited by the composition to responses elicited by a different composition.
- various assays can be utilized in order to characterize an antigen or composition and/or to determine whether an immune response has been stimulated in a T cell or group of T cells.
- assays are used to characterize a T cell response in a subject that has been administered an immunogenic composition to elicit an anti-chlamydia response (e.g., to evaluate whether a detectable T cell response has been elicited and/or to evaluate the potency of the response).
- the novel chlamydia antigens described herein also provide diagnostic agents to evaluate exposure to chlamydia infections (e.g., in non-vaccinated subjects).
- assays are used to characterize a T cell response in a subject to determine whether the subject has been infected with a chlamydia organism.
- the subject can be a subject suspected of exposure to a chlamydia organism recently (i.e., an assay to detect a response can be performed with a sample taken from the subject about 3, 4, 5, 6, 7, 8, 9, 10, 14, 30, or more days after suspected exposure to a chlamydia organism).
- the subject can be a subject suspected of exposure to a chlamydia organism weeks, months, or years prior to the assay.
- novel chlamydia antigens described herein also provide prognostic agents to evaluate outcomes of exposure to a chlamydia organism (e.g., in subjects known to be, or to have been, infected with a chlamydia organism).
- assays are used to characterize a T cell response in a subject to assess the likelihood of sequelae (e.g, pelvic inflammatory disease and infertility) to infection with a chlamydia organism.
- stimulation of an immune response in T cells is determined by measuring antigen-induced production of cytokines by T cells.
- stimulation of an immune response in T cells can be determined by measuring antigen-induced production of IFN- ⁇ , IL-4, IL-2, IL-6, IL-10, IL-17 and/or TNF-a by T cells.
- antigen-induced production of cytokines by T cells can be measured by intracellular cytokine staining followed by flow cytometry. Other suitable methods include surface capture staining followed by flow cytometry, or methods that determine cytokine concentration in supernatants of activated T cell cultures, such as ELISA or
- antigen-produced production of cytokines by T cells is measured by ELISPOT assay.
- ELISPOT assays typically employ a technique very similar to the sandwich enzyme-linked immunosorbent assay (ELISA) technique.
- An antibody e.g. monoclonal antibody, polyclonal antibody, etc. is coated aseptically onto a PVDF
- biotinylated polyclonal antibody specific for the cytokine is added to the wells. This antibody is reactive with a distinct epitope of the target cytokine and thus is employed to detect the captured cytokine. Following a wash to remove any unbound biotinylated antibody, the detected cytokine is then visualized using an avidin-HRP, and a precipitating substrate (e.g., AEC, BCIP/NBT).
- the colored end product (a spot, usually red or blue) typically represents an individual cytokine-producing cell. Spots can be counted manually (e.g., with a dissecting microscope) or using an automated reader to capture the microwell images and to analyze spot number and size. In some embodiments, each spot correlates to a single cytokine-producing cell.
- an immune response in T cells is said to be stimulated if between about 1% and about 100% of antigen- specific T cells produce cytokines. In some embodiments, an immune response in T cells is said to be stimulated if at least about 1%, at least about 5%, at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, at least about 99%, or about 100% of antigen- specific T cells produce cytokines.
- an immune response in T cells is said to be stimulated if immunized subjects comprise at least about 10-fold, at least about 50-fold, at least about 100-fold, at least about 500-fold, at least about 1000-fold, at least about 5000-fold, at least about 10,000-fold, at least about 50,000-fold, at least about 100,000-fold, or greater than at least about 100,000-fold more cytokine-producing cells than do naive controls.
- stimulation of an immune response in T cells can be determined by measuring antigen-induced proliferation of T cells.
- antigen-induced proliferation may be measured as uptake of H -thymidine in dividing T cells (sometimes referred to as "lymphocyte transformation test, or "LIT").
- LIT lymphocyte transformation test
- antigen-induced proliferation is said to have occurred if H-thymidine uptake (given as number of counts from a ⁇ counter) is at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 50-fold, at least about 100-fold, at least about 500-fold, at least about 1000-fold, at least about 5000-fold, at least about 10,000-fold, or greater than at least about 10,000-fold higher than a naive control.
- antigen-induced proliferation may be measured by flow cytometry.
- antigen-induced proliferation may be measured by a carboxyfluorescein succinimidyl ester (CFSE) dilution assay.
- CFSE is a non-toxic, fluorescent, membrane -permeating dye that binds the amino groups of cytoplasmic proteins with its succinimidyl-reactive group (e.g., T cell proteins). When cells divide, CFSE-labeled proteins are equally distributed between the daughter cells, thus halving cell fluorescence with each division.
- antigen-specific T cells lose their fluorescence after culture in the presence of the respective antigen (CFSE low ) and are distinguishable from other cells in culture (CFSE high ).
- antigen-induced proliferation is said to have occurred if CFSE dilution (given as the percentage of CFSE low cells out of all CFSE + cells) is at least about 5%, at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, or at least about 100%.
- an immune response in T-cells is said to be stimulated if cellular markers of T cell activation are expressed at different levels (e.g., higher or lower levels) relative to unstimulated cells.
- CD1 la, CD27, CD25, CD40L, CD44, CD45RO, and/or CD69 are more highly expressed in activated T cells than in unstimulated T cells.
- L-selectin (CD62L), CD45RA, and/or CCR7 are less highly expressed in activated T cells than in unstimulated T cells.
- an immune response in T cells is measured by assaying cytotoxicity by effector CD8 + T cells against antigen-pulsed target cells.
- a 51 chromium ( 51 Cr) release assay can be performed.
- effector CD8 + T cells bind infected cells presenting virus peptide on class I MHC and signal the infected cells to undergo apoptosis. If the cells are labeled with 51 Cr before the effector CD8 + T cells are added, the amount of 51 Cr released into the supernatant is proportional to the number of targets killed.
- an immune response in T cells is measured by an in vivo cytotoxicity assay in which target cells are antigen pulsed and labeled with a fluorescent dye, then transferred into immunized animals.
- Specific cytolytic T cells cause the disappearance of fluorescently labeled cells that are pulsed with a relevant antigen, but no decrease in cells pulsed with a control antigen.
- an immune response in T cells is measured by detecting expression of one or more of Perforin, Granzyme B, or CD107a (e.g., by ELISPOT or flow cytometry). See, e.g., Betts et al., J. Immunol. Meth. 281(l-2):65-78, 2003.
- various assays can be utilized in order to determine whether an immune response has been stimulated in a B cell or group of B cells, e.g., to characterize an antibody response in a subject that has been administered an immunogenic composition against chlamydia, or to determine whether a subject has been exposed to a chlamydia organism.
- stimulation of an immune response in B cells can be determined by measuring antibody titers.
- antibody titer refers to the ability of antibodies to bind antigens at particular dilutions.
- a high antibody titer refers to the ability of antibodies to bind antigens even at high dilutions.
- an immune response in B cells is said to be stimulated if antibody titers are measured to be positive at dilutions at least about 5-fold greater, at least about 10-fold greater, at least about 20-fold greater, at least about 50-fold greater, at least about 100-fold greater, at least about 500-fold greater, at least about 1000 fold greater, or more than about 1000-fold greater than in non-immunized individuals or pre-immune serum.
- stimulation of an immune response in B cells can be determined by measuring antibody affinity.
- an immune response in B cells is said to be stimulated if an antibody that has an equilibrium dissociation constant (K ⁇ j) less than 10 "7 M, less than 10 "8 M, less than 10 "9 M, less than 10 "10 M, less than 10 "11 M, less than
- K ⁇ j equilibrium dissociation constant
- a T cell-dependent immune response in B cells is said to be stimulated if class-switch recombination has occurred.
- a switch from IgM to another isotype e.g., to an IgG isotype or to IgA or to a mixture of these isotypes
- IgM is indicative of a T-cell dependent immune response in B cells.
- an immune response in B cells is determined by measuring affinity maturation of antigen- specific antibodies. Affinity maturation occurs during the germinal center reaction whereby activated B cells repeatedly mutate a region of the immunoglobulin gene that encodes the antigen-binding region. B cells producing mutated antibodies which have a higher affinity for antigen are preferentially allowed to survive and proliferate. Thus, over time, the antibodies made by B cells in GCs acquire incrementally higher affinities.
- the readout of this process is the presence of high antibody titer (e.g. high affinity IgG antibodies that bind and neutralize antigens even at high dilutions).
- an immune response in B cells is said to be stimulated if memory B cells and/or long-lived plasma cells that can produce large amounts of high- affinity antibodies for extended periods of time have formed.
- antibody titers are measured after different time intervals (e.g. 2 weeks, 1 month, 2 months, 6 months, 1 year, 2 years, 5 years, 10 years, 15 years, 20 years, 25 years, or longer) after vaccination in order to test for the presence of memory B cells and/or long-lived plasma cells that can produce large amounts of high-affinity antibodies for extended periods of time.
- memory B cells and/or long-lived plasma cells that can produce large amounts of high-affinity antibodies for extended periods of time are said to be present by measuring humoral responses (e.g. if humoral responses are markedly more rapid and result in higher titers after a later booster vaccination than during the initial sensitization).
- an immune response in B cells is said to be stimulated if a vigorous germinal center reaction occurs.
- a vigorous germinal center reaction can be assessed visually by performing histology experiments.
- vigorous germinal center reaction can be assayed by performing
- immunohistochemistry of antigen-containing lymphoid tissues e.g., vaccine-draining lymph nodes, spleen, etc.
- immunohistochemistry is followed by flow cytometry.
- stimulation of an immune response in B cells can be determined by identifying antibody isotypes (e.g., IgG, IgA, IgE, IgM).
- antibody isotypes e.g., IgG, IgA, IgE, IgM.
- production of IgG isotype antibodies by B cells is a desirable immune response by B cells.
- production of IgA isotype antibodies by B cells is a desirable immune response by B cells.
- an immune response in B cells is determined by analyzing antibody function in neutralization assays.
- the ability of a chlamydia organism to infect a susceptible cell in vitro in the absence of serum is compared to conditions when different dilutions of immune and non-immune serum are added to the culture medium in which the cells are grown.
- an immune response in a B cell is said to be stimulated if infection by a chlamydia organism is neutralized at a dilution of about 1:5, about 1: 10, about 1:50, about 1:100, about 1:500, about 1: 1000, about 1:5000, about 1: 10,000, or less.
- an immunogenic composition may be characterized
- B cell responses are characterized by transferring serum from immune mice as a "passive vaccine" to assess protection of non-immune mice from pathological effects or burden of infection.
- infiltrating leukocyte populations are characterized (e.g., to assess the number and type cells in a region of infection, e.g., whether CD4 + T cells, CD8 + T cells, or other cell types are present).
- Animal models for chlamydial urogenital infection have been described.
- a chlamydia organism is applied as an intravaginal inoculum, and infection and pathology of one or more of lower and upper genital tracts of the infected animal is characterized. See, e.g., Barron et al. (J. Infect. Dis.
- an immunogenic composition is assessed in an animal model of chlamydia infection.
- lower urogenital tract infection by chlamydia is assessed in the model (e.g., lower tract bacterial burden and/or inflammation due to infection is assessed).
- upper tract infection by chlamydia is assessed in the model (e.g., one or more of upper tract bacterial burden, inflammation, infertility, collagen deposition, scarring due to infection, are assessed).
- an ability to prevent ascension of a chlamydia infection from the lower tract to the upper genital tract is assessed.
- rate of bacterial clearance from the lower tract is assessed.
- rate of bacterial clearance from the upper tract is assessed.
- an immunogenic composition is assessed in an animal model in multiple strains of the animal of interest (e.g., multiple mouse strains).
- presence and size of hydrosalpinx is assessed.
- desirable immunogenic compositions are characterized as having one or more of the above effects in vivo (e.g., in an animal model).
- an immunogenic composition reduces lower urogenital tract infection by chlamydia bacteria.
- an immunogenic composition reduces lower tract bacterial burden.
- an immunogenic composition reduces lower tract inflammation due to infection.
- an immunogenic composition reduces upper tract infection by chlamydia.
- an immunogenic composition reduces one or more of upper tract bacterial burden, inflammation, infertility, collagen deposition, scarring due to a chlamydia infection.
- an immunogenic composition reduces ascension of a chlamydia infection from the lower tract to the upper genital tract. In some embodiments, an immunogenic composition increases the rate of bacterial clearance from the lower tract and/or the upper tract. In some embodiments, an immunogenic composition reduces presence and/or size of hydrosalpinx or salpyngitis due to infection. In some embodiments, an immunogenic composition has one or more of the above effects in multiple animal strains (e.g., multiple mouse strains).
- compositions and methods described herein can be used for the prophylaxis and/or treatment of any chlamydia infection, chlamydial disease, disorder, and/or condition.
- prophylaxis refers to uses before onset of symptoms due to a chlamydia infection, chlamydial disease, disorder, and/or condition and/or before known exposure to a chlamydia organism.
- Subjects include, but are not limited to, humans and/or other primates; and other animals susceptible to infection by chlamydia organisms, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and/or dogs; and/or birds, including commercially relevant birds such as chickens, ducks, geese, and/or turkeys.
- immunogenic compositions in accordance with the present invention may be used to treat, alleviate, ameliorate, relieve, delay onset of, inhibit progression of, reduce risk of infection by, and reduce severity of, and/or reduce incidence of one or more symptoms or features of a chlamydial disease, disorder, and/or condition.
- inventive an immunogenic composition may be used to treat, alleviate, ameliorate, relieve, delay onset of, inhibit progression of, reduce severity of, and/or reduce incidence of one or more symptoms or features of chlamydial infection (e.g., C. trachomatis infection, C. pneumoniae infection, C. psittaci infection).
- a method for the prophylaxis and/or treatment of chlamydia infection comprises administering a therapeutically effective amount of an immunogenic composition described herein to a subject in need thereof, in such amounts and for such time as is necessary to achieve the desired result.
- a "therapeutically effective amount" of an inventive immunogenic composition is that amount effective for reducing risk of infection by, or treating, alleviating, ameliorating, relieving, delaying onset of, inhibiting progression of, reducing severity of, and/or reducing incidence of one or more symptoms or features of chlamydia infection.
- a therapeutically effective amount may be determined on a population basis, and is not required to be an amount that naturally induces a protective response in a particular subject.
- inventive prophylactic and/or therapeutic protocols involve administering a therapeutically effective amount of one or more inventive immunogenic compositions to a healthy subject (i.e., a subject who does not display any symptoms of chlamydia infection and/or who has not been diagnosed with chlamydia infection).
- a healthy subject i.e., a subject who does not display any symptoms of chlamydia infection and/or who has not been diagnosed with chlamydia infection.
- healthy individuals may be vaccinated using inventive
- immunogenic compositions prior to development of chlamydia infection and/or onset of symptoms of chlamydia infection can be treated substantially contemporaneously with (e.g., within 48 hours, within 24 hours, or within 12 hours of) the onset of symptoms of and/or exposure to chlamydia infection.
- individuals known to have chlamydia infection may receive treatment at any time.
- inventive prophylactic and/or therapeutic protocols involve administering a therapeutically effective amount of one or more inventive immunogenic compositions to a subject such that an immune response is stimulated in both T cells and B cells.
- immune responses e.g. T cell and/or B cell responses
- immune responses can be tailored to preferentially elicit the most desirable type of immune response for a given indication, e.g., humoral response, Thl T cell response, Thl7 T cell response, IFN- ⁇ secretion by antigen- specific T cells, cytotoxic T cell response, antibody response, B cell response, innate immune response, or a combination of these responses.
- Immunogenic Compositions e.g. T cell and/or B cell responses
- the present invention provides immunogenic compositions (e.g., vaccines) comprising a novel chlamydia antigen, e.g., one or more of a polypeptide antigen selected from Table 1, Table 2, Table 3, or combinations thereof, and one or more pharmaceutically acceptable excipients.
- a method of administering an inventive immunogenic composition to a subject in need thereof is provided.
- inventive compositions are administered to humans.
- the phrase "active ingredient" generally refers to an inventive
- immunogenic composition comprising at least one chlamydia antigen and optionally comprising one or more additional agents, such as an adjuvant.
- immunogenic compositions are principally directed to compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of immunogenic compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and/or perform such modification with merely ordinary, if any, experimentation.
- Subjects to which administration of the immunogenic compositions of the invention is contemplated include, but are not limited to, humans and/or other primates; mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, and/or dogs; and/or birds, including commercially relevant birds such as chickens, ducks, geese, and/or turkeys.
- the formulations of the immunogenic compositions described herein may be prepared by any method known or hereafter developed in the art of vaccines.
- such preparatory methods include the step of bringing the antigen(s) (or nucleic acids encoding the antigens, for nucleic acid based applications) into association with one or more excipients and/or one or more other accessory ingredients, and then, if necessary and/or desirable, shaping and/or packaging the product into a desired single- or multi-dose unit.
- An immunogenic composition of the invention may be prepared, packaged, and/or sold in bulk, as a single unit dose, and/or as a plurality of single unit doses.
- a "unit dose" is discrete amount of the immunogenic composition comprising a predetermined amount of the antigen(s).
- compositions of the invention will vary, depending upon the identity, size, and/or condition of the subject treated and further depending upon the route by which the composition is to be administered.
- Immunogenic formulations of the present invention may additionally comprise a pharmaceutically acceptable excipient, which, as used herein, includes any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.
- a pharmaceutically acceptable excipient includes any and all solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like, as suited to the particular dosage form desired.
- the pharmaceutically acceptable excipient is at least
- the excipient is approved for use in humans and for veterinary use. In some embodiments, the excipient is approved by United States Food and Drug Administration. In some embodiments, the excipient is pharmaceutical grade. In some embodiments, the excipient meets the standards of the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), the British Pharmacopoeia, and/or the International Pharmacopoeia.
- USP United States Pharmacopoeia
- EP European Pharmacopoeia
- British Pharmacopoeia the British Pharmacopoeia
- International Pharmacopoeia International Pharmacopoeia
- immunogenic compositions include, but are not limited to, inert diluents, dispersing and/or granulating agents, surface active agents and/or emulsifiers, disintegrating agents, binding agents, preservatives, buffering agents, lubricating agents, and/or oils. Such excipients may optionally be included in the inventive formulations.
- injectable formulations for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents.
- a sterile injectable preparation may be a sterile injectable solution, suspension or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
- acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S. P. and isotonic sodium chloride solution.
- sterile, fixed oils are conventionally employed as a solvent or suspending medium.
- any bland fixed oil can be employed including synthetic mono- or diglycerides.
- fatty acids such as oleic acid are used in the preparation of injectables.
- Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
- an immunogenic composition is administered to a mucosal surface.
- Compositions for rectal or vaginal administration can include suppositories which can be prepared by mixing immunogenic compositions of this invention with suitable excipients such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release antigen.
- an immunogenic composition is administered orally.
- Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules.
- the antigen can be mixed with at least one inert, pharmaceutically acceptable excipient such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monoste
- Suitable devices for use in delivering immunogenic compositions by an intradermal route described herein include short needle devices such as those described in U.S. Patents 4,886,499; 5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662. Jet injection devices which deliver liquid immunogenic compositions to the dermis via a liquid jet injector and/or via a needle which pierces the stratum corneum and produces a jet which reaches the dermis are suitable. Jet injection devices are described, for example, in U.S.
- Ballistic powder/particle delivery devices which use compressed gas to accelerate an immunogenic composition in powder form through the outer layers of the skin to the dermis are suitable.
- conventional syringes may be used in the classical mantoux method of intradermal administration.
- a therapeutically effective amount of an inventive immunogenic composition is delivered to a patient and/or animal prior to, simultaneously with, and/or after exposure to a chlamydia organism or diagnosis with a chlamydial disease, disorder, and/or condition.
- a therapeutic amount of an inventive composition is delivered to a patient and/or animal prior to, simultaneously with, and/or after onset of symptoms of a chlamydial disease, disorder, and/or condition.
- the amount of an immunogenic composition is sufficient to reduce risk of infection by, or treat, alleviate, ameliorate, relieve, delay onset of, inhibit progression of, reduce severity of, and/or reduce incidence of one or more symptoms or features of the chlamydial disease, disorder, and/or condition.
- Immunogenic compositions may be administered using any amount and any route of administration effective for treatment.
- the exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the particular composition, its mode of administration, its mode of activity, and the like.
- the specific effective dose level for any particular subject or organism will depend upon a variety of factors including the immunogenicity of the antigen composition employed; the specific composition employed; the nature of adjuvant used; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and like factors well known in the medical arts.
- Immunogenic compositions of the present invention may be administered by any route that elicits an immune response.
- an immunogenic composition is administered subcutaneously.
- an immunogenic composition is administered intramuscularly.
- the immunogenic compositions of the present invention are administered by a variety of routes, including oral, intravenous, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal, interdermal, rectal, intravaginal, intraperitoneal, topical (as by powders, ointments, creams, and/or drops), transdermal, mucosal, nasal, buccal, enteral, sublingual; by intratracheal instillation, bronchial instillation, and/or inhalation; and/or as an oral spray, nasal spray, and/or aerosol.
- an immunogenic composition of the invention may be administered in amounts that include a protein antigen in ranges of 1 ⁇ g-500 ⁇ g. In some embodiments, a dose of about 10 ⁇ g, 20 ⁇ g, 30 ⁇ g, 50 ⁇ g, or 100 ⁇ g is administered to a human.
- an immunogenic composition is administered more than once (e.g., twice, three times, four times, five times).
- a boost is given about one week, two weeks, three weeks, one month, three months, six months, one vear. or lonser after an initial immunization. Kits
- kits comprising one or more of the antigens described herein.
- the invention provides a kit including a novel chlamydia antigen and instructions for use.
- a kit may include multiple different chlamydia antigens.
- a kit may include any of a number of additional components or reagents in any combination. All of the various combinations are not set forth explicitly but each
- a kit may include, for example, (i) an immunogenic composition including at least one of the following chlamydia antigens: CT062, CT572, CT043, CT570, CT177, CT725, CT067, CT476, p6, CT310, or CT638 polypeptide antigens; and (ii) instructions for administering the composition to a subject in need thereof.
- the kit further includes an adjuvant.
- Kits that include nucleic acids encoding chlamydia antigens are also provided.
- a kit may include, for example, (i) a composition including a nucleic acid encoding a chlamydia antigen; (ii) instructions for use of the nucleic acid compositin (e.g., instructions for expressing the nucleic acid for producing the antigen, or instructions for administering the composition to a subject in need thereof to elicit a response against chlamydia).
- kits may, for example, include protocols and/or describe conditions for production of immunogenic compositions and/or administration of immunogenic compositions, to a subject in need thereof, etc.
- Kits generally include one or more vessels or containers so that some or all of the individual components and reagents may be separately housed.
- Kits may also include a means for enclosing individual containers in relatively close confinement for commercial sale, e.g., a plastic box, in which instructions, packaging materials such as styrofoam, etc., may be enclosed.
- An identifier e.g., a bar code, radio frequency identification (ID) tag, etc., may be present in or on the kit or in or one or more of the vessels or containers included in the kit.
- An identifier can be used, e.g., to uniquely identify the kit for purposes of quality control, inventory control, tracking, movement between workstations, etc. EXEMPLIFICATION
- Example 1 Peripheral blood mononuclear cells and plasma from women with a clinical history of Chlamydia trachomatis infection are used to identify chlamydia protein antigens
- PBMC peripheral blood mononuclear cells
- T cells were enriched from PBMC by magnetic bead depletion using the Miltenyi Pan T sorting kit following the manufacturer's instructions. The resulting enriched T cell population was then sorted using antibody-conjugated magnetic beads specific for CD4 + T cells (Miltenyi). The CD4 negative population was considered to be CD8 + . (In some cases, the PBMC depleted of T cells were cyropreserved.) Both T cell subsets were non- specifically expanded in vitro using magnetic beads coated with anti-CD3 and anti-CD28 antibodies (Dynal T Cell Expander).
- the T cells were maintained at 10 6 cells/mL in AIM-V- 5% (AIM-V, 5% FCS, Non-essential Amino Acids, Sodium Pyruvate, L-Glutamine, and beta-mercaptoethanol) plus recombinant IL-2.
- AIM-V- 5% AIM-V, 5% FCS, Non-essential Amino Acids, Sodium Pyruvate, L-Glutamine, and beta-mercaptoethanol
- the CD3/CD28 magnetic beads were removed from culture, and the enriched and expanded CD4 + and CD8 + T cells were separately screened using a chlamydia ORFeome library to determine which antigens naturally induced T cell responses.
- T cell screening required the co-culture of expanded T cells with autologous antigen presenting cells (APC) that were pulsed with the proteomic library.
- APC autologous antigen presenting cells
- APC were pulsed with induced bacteria from the proteomic library at a 100: 1 ratio of induced bacteria to APC.
- Method 1 plated 10 4 MDCC per well in 384- well flat bottom plates.
- Method 2 plated 10 5 APC per well comprised of MDCC and thawed T cell-depleted PBMC in 96- well round bottom plates.
- screen plates containing APC and library-expressing bacteria were placed in a 37°C, 5% C0 2 humidified incubator. After a two-hour incubation, the APC were washed with PBS and then fixed with 1% paraformaldehyde (PFA).
- PFA paraformaldehyde
- T cells were washed extensively, then expanded T cells were added to the pulsed, fixed APC and the plates returned to a 37°C, 5% C0 2 humidified incubator.
- 4xl0 4 T cells were added to the 10 4 pulsed MDCC plated in each well of the 384- well plates described in Method 1.
- up to 10 5 T cells were added to the 10 5 pulsed APC plated in each well of the 96- well plates described in Method 2.
- T cell responses were monitored by measuring interferon gamma (IFN- ⁇ ) in the cell-free supernatants by ELISA (BD OptEIA kit).
- IFN- ⁇ interferon gamma
- FIG. 1, 2, and 3 depict exemplary graphs illustrating the frequency with which identified antigens were recognized by, respectively, CD4 + and CD8 + T cells obtained from protected and unprotected donors. Based on evaluation of negative controls, donor and plate variation, a donor was classified as a "responder” if the fold ratio of the value over negative control was greater than 1.63 (CD4 + ) or 1.66 (CD8 + ). Percent responders >10 indicated a higher number of responders than due to chance alone. Statistical significance was reached when the percent responders was >15 (all donors, including negative controls), or approximately 19% (protected and unprotected donors). FIG. 1 and FIG. 2 depict separate exemplary results for protected and unprotected donors. Four C.
- FIG. 3 depicts an exemplary result illustrating CD4 + , CD8 + , and combined T cell responses for all donors (protected and unprotected). Antigens represented at the highest overall frequency, whether or not represented at statistically higher frequency in protected donors, are also attractive candidates for vaccine, diagnostic and prognostic applications.
- the plates were then sealed and shipped for printing onto microarrays (Gentel Biosceinces, Inc.).
- the protein microarrays were probed with plasma samples of subjects recruited for T cell screens above.
- An antibody specific for human IgG was used to probe the bound plasma samples for protein specific antibody and detected by chromogenic substrate. Responses were considered positive if the signal was statistically significantly above the background value of negative controls.
- Two criteria were used for selection: the first was overall frequency of responses across all cohorts and the second was responses with statistically greater frequency in protected subjects as compared to unprotected donors, with a p-value of ⁇ 0.05.
- FIG. 4 depicts an exemplary result illustrating the frequency with which chlamydia antigens were bound by IgG present in donor sera, i.e. have elicited a donor B cell response.
- the left side of the panel displays chlamydia antigens detected by IgG with overall highest frequency across all donors (protected and unprotected).
- the right side of the panel displays chlamydia antigens detected by IgG with statistically greater frequency in protected donors as compared to unprotected donors.
- Example 2 Identified chlamydia protein antigens are immunogenic in mice
- mice were immunized subcutaneously in the scruff of the neck with a 100 ⁇ injection of 5 ⁇ g antigen plus adjuvant (12 ⁇ g dose of an ISCOM matrix with a 91:9 mixture of Quillaja saponin matrix A and matrix C) in saline. The mice received two injections, 21 days apart. Seven days after the final injection, mice were euthanized, and blood and tissues harvested for further analysis.
- CD8 + T cells were enriched from mouse splenocytes using magnetic beads, starting from mouse spleens harvested above.
- Membrane plates were prepared by coating overnight with capture antibody specific for IFN- ⁇ and subsequently blocked with supplemented medium for a minimum of 2 hours at 37°C.
- APCs were prepared by pulsing naive T-depleted splenocytes with antigen for 2 hours at 37°C.
- CD4 + ELISPOTs APCs were pulsed with whole protein.
- CD8 + ELISPOTs ISCOM matrix at a concentration of 20 ⁇ g/mL was added to the whole protein to facilitate antigen uptake and processing.
- the APCs and T cells were added to appropriate wells of the pre-coated plates.
- a negative control was APCs incubated for 2 hours at 37°C with no additional antigen, and a positive control was T cells incubated with phorbol myristate acetate (PMA) and ionomycin. The plates were allowed to incubate for 18 hours at 37°C under 5% C0 2 . The spots were visualized using a secondary
- biotinylated antibody specific for IFN- ⁇ horseradish peroxidase (HRP) and 3-amino-9- ethylcarbazole (AEC) substrate.
- HRP horseradish peroxidase
- AEC 3-amino-9- ethylcarbazole
- FIG. 5 depicts an exemplary result illustrating IFN- ⁇ levels induced ex vivo in
- FIG. 5A depicts an exemplary result illustrating antigens that were originally identified through T cell responses.
- FIG. 5B depicts an exemplary result illustrating antigens that were originally identified through B cell responses, demonstrating that these antigens can in some cases also elicit robust T cell responses.
- Antigen-specific serum antibody titers of immunized mice were determined by direct protein ELISA. Blood was collected 7 days post last injection by terminal cardiac puncture. The sera were processed and stored at -80°C. ELISA plates were coated overnight at 4°C with 5 ⁇ g of whole protein in 0.1 M carbonate buffer, pH 9.5. Plates were washed with TBS + 0.05% Tween-20 (TBS-T) and blocked with TBS-T + 1% bovine serum albumin for lh. Serum samples were serially diluted and incubated in the antigen-coated wells for 2 hours at room temperature.
- AP activity was achieved by the addition of p-Nitrophenyl phosphate (pNPP; Sigmafast, Sigma- Aldrich), and the reaction stopped with 3N NaOH and absorbance read at 405 nm.
- Endpoint titers were calculated by extrapolation of the linear portion of the serial dilutions and defining the endpoint as the dilution at which the linear portion of the curve intersects with the background cut-off. The cut-off used for data calculation was 2 times the value of the negative control serum from a naive mouse.
- FIG. 6 depicts an exemplary result illustrating IgG antibody titers against the indicated chlamydia antigens, following immunization with the same antigen. Results shown in the left side of the panel demonstrate that antigens originally identified through T cell responses (e.g. FIG. 1, 2 and 3) can in some cases also elicit robust B cell responses.
- Example 3 Mice immunized with identified chlamydia protein antigens are protected against Chlamydia trachomatis challenge
- mice C57BL/6 mice (8 per group) were immunized subcutaneously in the scruff of the neck with a 100 ⁇ injection of 5 ⁇ g antigen plus adjuvant (24 ⁇ g dose of an ISCOM matrix with a 91:9 mixture of Quillaja saponin matrix A and matrix C) in saline. The mice received two injections, 21 days apart. Depo-Provera (1.25 mg) was administered
- McCoy cells, and elementary bodies were purified by RenoCal-76 gradient centrifugation and stored in sucrose phosphate (SPG) buffer.
- SPG sucrose phosphate
- the mice were challenged seven days after the last immunization by intravaginal deposition of 0.5-1 xlO 6 IFU Chlamydia trachomatis serovar D elementary bodies directly onto the ectocervix with a positive displacement pipet.
- FIG. 7 depicts an exemplary result illustrating reduction of ectocervical chlamydia burden in mice immunized with the indicated chlamydia protein antigens and subsequently intravaginally infected with Chlamydia trachomatis.
- FIG. 7A depicts an exemplary result for representative chlamydia protein antigens CT062, CT043, and for the combination CT062 + CT043.
- FIG. 7B depicts an exemplary result for representative chlamydia protein antigen combination CT638 + CT476.
- Oviducts and ovaries were collected from immunized and challenged mice at day 21 post-infection. Chlamydia, living and dead, present in whole oviducts and ovaries were detected by real-time quantitative PCR. The oviducts and ovaries were digested overnight at 56°C in tissue lysis buffer containing 0.6mg Proteinase K. DNA was extracted using the QIAamp DNA Mini Kit (Qiagen) according to manufacturer's instructions.
- Extracted DNA was subjected to PCR with primers specific for Chlamydia trachomatis 16SrRNA gene. Briefly, 15 ⁇ of extracted DNA was processed in a 20uL reaction volume containing 0.8uM of each primer and 1 U of Taq polymerase. Amplifications were carried out in a StepOnePlus Real-Time PCR system (Applied Biosystems). The gene copy number was determined by extrapolation using a standard curve of Chlamydia 16s rRNA purified plasmid of known copy number.
- FIG. 8 depicts an exemplary result illustrating reduction of upper reproductive tract chlamydia burden in mice immunized with the indicated chlamydia protein antigens and subsequently intravaginally infected with Chlamydia trachomatis.
- FIG. 8A depicts an exemplary result for representative chlamydia protein antigens CT062, CT043, and for the combination CT062 + CT043.
- UVEB indicates responses from mice immunized with the positive control, UV-inactivated whole Chlamydia trachomatis elementary bodies.
- FIG. 8B depicts an exemplary result for representative chlamydia protein antigens CT067, CT0788tm, and CT328.
- Example 4 Subsequent to infection with Chlamydia trachomatis, lymphatic and splenic T cells are primed to respond to identified chlamydia protein antigens.
- mice were intravaginally infected with lxlO 6 IFU purified
- FIG. 9 depicts an exemplary result illustrating induction of IFN- ⁇ in CD4 +
- MCVSRSLRWCLCFLLLCGWVDAGVYDKLRLTGI I IDRNGLSETICSKEKLQKYTKIDFLSPQPYQKVMRTYKNA AGESVACLTTYYPNGQIRQYLECLNNRAFGRYREWHSNGKIHIQAEVIGGIADLHPSAEAGWLFDGTTYAHDSEG RLEAVIHYEKGLLEGI SLYYHANGNVWKECPYHKGVAHGDFLVFTEEGSLLKKQTFCKGQLSGCVLRYEPGSQSL LSEEEYKQGKLRSGKYYDPLTKEEIACVVNGKGKQVIYGKYAI IETRQIVHGVPHGEVLLFDEHGKSLLQAYSLI NGQKEGEEVFFYPGGEGRKMLLTWSQGILQGAVKTWYPNGALESSKELVQNKKTGILMLYYPEGQVMATEEYVDD LLIKGEYFRPNDRYPYAKVEKGCGTAVFFSATGGLLKKVLYEDG
- the invention includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process. Furthermore, it is to be understood that the invention encompasses all variations, combinations, and permutations in which one or more limitations, elements, clauses, descriptive terms, etc., from one or more of the listed claims is introduced into another claim. For example, any claim that is dependent on another claim can be modified to include one or more limitations found in any other claim that is dependent on the same base claim.
- any particular embodiment of the present invention that falls within the prior art may be explicitly excluded from any one or more of the claims. Since such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the compositions of the invention (e.g., any antigen, any method of administration, any prophylactic and/or therapeutic application, etc.) can be excluded from any one or more claims, for any reason, whether or not related to the existence of prior art.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Immunology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Urology & Nephrology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Endocrinology (AREA)
- Reproductive Health (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CA2849391A CA2849391A1 (en) | 2010-10-20 | 2011-10-20 | Chlamydia antigens and uses thereof |
AU2011316924A AU2011316924A1 (en) | 2010-10-20 | 2011-10-20 | Chlamydia antigens and uses thereof |
EP11835162.6A EP2629793A4 (en) | 2010-10-20 | 2011-10-20 | Chlamydia antigens and uses thereof |
JP2013535100A JP2013545448A (en) | 2010-10-20 | 2011-10-20 | Chlamydia antigen and uses thereof |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US40516210P | 2010-10-20 | 2010-10-20 | |
US61/405,162 | 2010-10-20 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2012054755A1 true WO2012054755A1 (en) | 2012-04-26 |
Family
ID=45975625
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2011/057143 WO2012054755A1 (en) | 2010-10-20 | 2011-10-20 | Chlamydia antigens and uses thereof |
Country Status (6)
Country | Link |
---|---|
US (2) | US20120135025A1 (en) |
EP (1) | EP2629793A4 (en) |
JP (2) | JP2013545448A (en) |
AU (2) | AU2011316924A1 (en) |
CA (1) | CA2849391A1 (en) |
WO (1) | WO2012054755A1 (en) |
Families Citing this family (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3590533A1 (en) | 2009-05-22 | 2020-01-08 | Genocea Biosciences, Inc. | Vaccines against herpes simplex virus type 2: compositions and methods for eliciting an immune response |
JP2013525271A (en) | 2010-03-12 | 2013-06-20 | チルドレンズ メディカル センター コーポレーション | Immunogen and screening method thereof |
EP2643014A4 (en) | 2010-11-24 | 2015-11-11 | Genocea Biosciences Inc | Vaccines against herpes simplex virus type 2: compositions and methods for eliciting an immune response |
CA2885693C (en) * | 2011-11-23 | 2020-07-28 | Genocea Biosciences, Inc. | Nucleic acid vaccines against herpes simplex virus type 2: compositions and methods for eliciting an immune response |
WO2014124228A1 (en) | 2013-02-07 | 2014-08-14 | Children's Medical Center Corporation | Protein antigens that provide protection against pneumococcal colonization and/or disease |
WO2018064232A1 (en) | 2016-09-28 | 2018-04-05 | Genocea Biosciences, Inc. | Methods and compositions for treating herpes |
EP3506625B1 (en) | 2016-10-14 | 2021-06-09 | Panasonic Intellectual Property Management Co., Ltd. | Projection system, projection method, flying object system, and flying object |
WO2020056202A1 (en) | 2018-09-12 | 2020-03-19 | Affinivax, Inc. | Multivalent pneumococcal vaccines |
AU2022342080A1 (en) | 2021-09-09 | 2024-03-28 | Affinivax, Inc. | Multivalent pneumococcal vaccines |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080317772A1 (en) * | 1998-12-08 | 2008-12-25 | Corixa Corporation | Compounds and methods for treatment and diagnosis of chlamydial infection |
US20090304722A1 (en) * | 2004-10-25 | 2009-12-10 | Michael Theisen | Chlamydia trachomatis antigens for vaccine and diagnostic use |
WO2010078027A1 (en) * | 2008-12-17 | 2010-07-08 | Genocea Biosciences, Inc. | Chlamydia antigens and uses thereof |
Family Cites Families (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7041490B1 (en) * | 1997-11-28 | 2006-05-09 | Serono Genetics Institute, S.A. | Chlamydia trachomatis polynucleotides and vectors, recombinant host cells, DNA chips or kits containing the same |
EP1133572A4 (en) * | 1998-11-12 | 2005-06-15 | Univ California | Chlamydia pneumoniae genome sequence |
NZ546711A (en) * | 2001-12-12 | 2008-06-30 | Chiron Srl | Immunisation against chlamydia trachomatis |
DE602004028705D1 (en) * | 2003-02-24 | 2010-09-30 | Pasteur Institut | APPLICATION OF THE CPn104 POLYPEPTIDE FROM CHLAMYDIA PNEUMONIAE OR ITS ANTIBODIES FOR THE DIAGNOSIS OF A CHLAMYDIA INFECTION |
MXPA05013260A (en) * | 2003-06-26 | 2006-03-09 | Chiron Corp | Immunogenic compositions for chlamydia trachomatis. |
GB0428394D0 (en) * | 2004-12-24 | 2005-02-02 | Chiron Srl | Saccharide conjugate vaccines |
AU2006341122B2 (en) * | 2005-12-22 | 2013-08-22 | Novartis Vaccines And Diagnostics, Srl. | Chlamydial antigens |
US9068007B2 (en) * | 2008-10-09 | 2015-06-30 | Board Of Regents, The University Of Texas System | Methods and compositions for chlamydial antigens for diagnosis and treatment of chlamydial infection and disease |
JP2012519482A (en) * | 2009-03-06 | 2012-08-30 | ノバルティス アーゲー | Chlamydia antigen |
-
2011
- 2011-10-20 JP JP2013535100A patent/JP2013545448A/en active Pending
- 2011-10-20 AU AU2011316924A patent/AU2011316924A1/en not_active Abandoned
- 2011-10-20 WO PCT/US2011/057143 patent/WO2012054755A1/en active Application Filing
- 2011-10-20 US US13/277,996 patent/US20120135025A1/en not_active Abandoned
- 2011-10-20 CA CA2849391A patent/CA2849391A1/en not_active Abandoned
- 2011-10-20 EP EP11835162.6A patent/EP2629793A4/en not_active Withdrawn
-
2016
- 2016-02-24 JP JP2016032672A patent/JP2016106117A/en active Pending
- 2016-03-24 US US15/080,324 patent/US20170043005A1/en not_active Abandoned
- 2016-04-21 AU AU2016202528A patent/AU2016202528A1/en not_active Abandoned
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20080317772A1 (en) * | 1998-12-08 | 2008-12-25 | Corixa Corporation | Compounds and methods for treatment and diagnosis of chlamydial infection |
US20090304722A1 (en) * | 2004-10-25 | 2009-12-10 | Michael Theisen | Chlamydia trachomatis antigens for vaccine and diagnostic use |
WO2010078027A1 (en) * | 2008-12-17 | 2010-07-08 | Genocea Biosciences, Inc. | Chlamydia antigens and uses thereof |
Non-Patent Citations (1)
Title |
---|
See also references of EP2629793A4 * |
Also Published As
Publication number | Publication date |
---|---|
JP2013545448A (en) | 2013-12-26 |
US20170043005A1 (en) | 2017-02-16 |
EP2629793A4 (en) | 2014-05-28 |
CA2849391A1 (en) | 2012-04-26 |
EP2629793A1 (en) | 2013-08-28 |
JP2016106117A (en) | 2016-06-16 |
US20120135025A1 (en) | 2012-05-31 |
AU2011316924A1 (en) | 2013-05-02 |
AU2016202528A1 (en) | 2016-05-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20170043005A1 (en) | Chlamydia antigens and uses thereof | |
US20110293664A1 (en) | Chlamydia antigens and uses thereof | |
JP4896715B2 (en) | Immunogenic composition against Chlamydiatrachomatis | |
JP6916828B2 (en) | A protein antigen that provides protection from pneumococcal colonization and / or disease | |
US8481057B2 (en) | Chlamydial antigens | |
JP2008544745A (en) | Immunogenic composition for Chlamydiatrachomatis | |
CN105377879B (en) | Vaccine against chlamydia species | |
JP2007526318A (en) | Immunogenic composition against Chlamydiapneumoniae | |
JP2012519482A (en) | Chlamydia antigen | |
JP2010505795A (en) | Vaccines against Chlamydia infection | |
US10835594B2 (en) | Type III secretion injectisome proteins for treatment and prevention of chlamydial infections | |
Shillova | Per-oral Inoculation with Live or Killed Chlamydia muridarum Protects against a Chlamydia muridarum Per-vaginal Challenge | |
MAHONY et al. | Patent 2973220 Summary | |
Murdin et al. | Vaccines for Chlamydia trachomatis and Chlamydia pneumoniae |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 11835162 Country of ref document: EP Kind code of ref document: A1 |
|
ENP | Entry into the national phase |
Ref document number: 2013535100 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
ENP | Entry into the national phase |
Ref document number: 2011316924 Country of ref document: AU Date of ref document: 20111020 Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2011835162 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2011835162 Country of ref document: EP |
|
ENP | Entry into the national phase |
Ref document number: 2849391 Country of ref document: CA |