Method for enhancing an immune response

• US 6,514,948 B1
• Filed: 07/02/1999
• Issued: 02/04/2003
• Est. Priority Date: 07/02/1999
• Status: Expired due to Fees

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Raz, Eyal R., Kobayashi, Hiroko
• Primary Examiner(s): Wehbe, Anne M.
• Assistant Examiner(s): LI, QIAN JANICE
• Application Number:
  US09/347,343
• Time in Patent Office: 1,313 Days
• Field of Search: 424/184.1, 424/234.1, 424/278.1, 424/282.1, 514/44, 536/23.1, 536/24.1, 536/24.5, 435/69.1, 435/320.1, 435/325, 435/455, 435/459
• US Class Current: 514/44.00R
• CPC Class Codes: A61K 2039/55561 CpG containing adjuvants; Oligonucleotide containing adjuvants
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 39/39 characterised by the immunostimulating additives, e.g. chemical adjuvants
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61P 37/04 Immunostimulants

Abstract

Disclosed is a method for enhancing an immune response to a substance, such as an antigen or microbial pathogen. The immune response can be, for example, production of IgG2 antibodies. The method comprises administering an immunostimulatory nucleotide sequence (ISS) to a subject at least one hour prior to exposure to the substance by the subject. The subject may be exposed to the substance either naturally, as with an environmental pathogen, or by administration, as with a known antigen. The method can be used for protecting or immunizing a subject against an antigen or pathogen, providing more effective immunization than if the ISS were co-administered with the substance. The method can be used prophylactically or therapeutically. In preferred embodiments, the ISS comprises a CG, p(GC) or p(IC) DNA or RNA nucleotide sequence. Of these, a CG containing nucleotide sequence is preferred. The ISS can further comprise a pG nucleotide sequence. Examples of an ISS include sequences comprising 5′-rrcgyy-3′, 5′-rycgyy-3′, 5′-rrcgyycg-3′ or 5′-rycgyycg-3′. The ISS is preferably administered between about 6 hours and about 6 weeks prior to exposure to the substance, and more preferably between about 1 day and about 4 weeks prior. Most preferably, the ISS is administered between about 3 days and about 8 days prior to exposure to the substance. The ISS can be administered via a mucosal or systemic route. The substance can be an antigen or pathogen associated with an infectious disease, an allergen or a cancer.

Claims

1. A method for enhancing an immune response to an antigenic substance, comprising administering intranasally a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS) to a subject antigenic about one day to about seven days prior to exposure to the antigenic substance by the subject, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the antigenic substance, an immune response specific for the antigenic substance antigenic is enhanced relative to an immune response produced by administration of the substance alone.

9. The method of any one of claims 1-8, wherein the antigenic substance is an antigen of an infectious pathogen, an allergen or a cancer antigen.

10. The method of any one of claims 1-8, wherein the antigenic substance is an antigen of an infectious pathogen selected from the group consisting of a virus, a bacterium, a mycobacterium, and a parasite.

11. The method of any one of claims 1-8, wherein the polynucleotide comprises a backbone phosphate group modification.

12. The method of any one of claims 1-8, wherein the polynucleotide comprises a backbone phosphate group modification comprising a phosphorothioate or a phosphorodithioate.

13. The method of any one of claims 1-8, wherein the immune response is a Th1 response.

14. The method of any one of claims 1-8, wherein the polynucleotide is a plasmid.

2. A method of immunizing a subject against an antigenic substance, comprising administering intranasally to the subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS) about one day to about seven days prior to exposing the subject to the antigenic substance, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the substance, an immune response specific for the antigenic substance is enhanced relative to an immune response produced by administration of the antigenic substance alone, and the subject is immunized against the antigenic substance.

3. A method of eliciting production of an antibody associated with a Th1 immune response, comprising administering intranasally to a subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS) from about one day to about seven days prior to administration of an antigenic substance to the subject, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rrcgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the antigenic substance, production of antibody associated with a Th1 immune response specific for the antigenic substance is enhanced relative to an immune response produced by administration of the antigenic substance alone.

4. A method for modulating an immune response in a subject, the method comprising:

administering intranasally to a subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS), wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rycgyy-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and wherein said polynucleotide is administered about one day to about seven days prior to exposure to an antigenic substance and in an amount effective to modulate an immune response specific for the antigenic substance in the subject upon exposure to the antigenic substance.

5. A method for modulating an immune response in a subject, the method comprising:

administering intradermally to a subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS), wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and wherein said polynucleotide is administered about three days to about 14 days prior to exposure to an antigenic substance and in an amount effective to modulate an immune response specific for the antigenic substance in the subject upon exposure to the antigenic substance.

6. A method for enhancing an immune response to an antigenic substance, comprising administering intradermally a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS) to a subject said administering being about three days to about 14 days prior to exposure to the substance by the subject, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the substance, an immune response to the substance is enhanced relative to an immune response produced by administration of the substance alone.

7. A method of immunizing a subject against an antigenic substance, comprising administering intradermally to the subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS), said administering being about three days to about 14 days prior to exposing the subject to the antigenic substance, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the antigenic substance, an immune response specific for the antigenic substance is enhanced relative to an immune response produced by administration of the antigenic substance alone, and the subject is immunized against the antigenic substance.

8. A method of eliciting production of an antibody associated with a Th1 immune response, comprising administering intradermally to a subject a polynucleotide comprising an immunostimulatory nucleotide sequence (ISS), said administering being about three days to about 14 days prior to administration of an antigenic substance to the subject, wherein the ISS comprises the sequence 5′

-rrcgyy-3′

(SEQ ID NO;

     1), 5′

-rycgyy-3′

(SEQ ID NO;

     2), 5′

-rrcgyycg-3′

(SEQ ID NO;

     3) or 5′

-rycgyycg-3′

(SEQ ID NO;

     4), and

wherein upon exposure to the antigenic substance, production of antibody associated with a Th1 immune response specific for the antigenic substance is enhanced relative to an immune response produced by administration of the antigenic substance alone.