Immunogenic composition of interleukin-12 (IL-12), alum,...

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Virus or component thereof

Reexamination Certificate

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C424S278100, C424S204100, C424S231100, C536S023720

Reexamination Certificate

active

06488936

ABSTRACT:

BACKGROUND OF THE INVENTION
The immune system uses many mechanisms for attacking pathogens; however, not all of these mechanisms are necessarily activated after immunization. Protective immunity induced by vaccination is dependent on the capacity of the vaccine to elicit the appropriate immune response to resist or eliminate the pathogen. Depending on the pathogen, this may require a cell-mediated and/or humoral immune response.
The current paradigm for the role of helper T cells in the immune response is that T cells can be separated into subsets on the basis of the cytokines they produce, and that the distinct cytokine profile observed in these cells determines their function. This T cell model Includes two major subsets: TH-1 cells that produce IL-2 and interferon &ggr; (IFN-&ggr;) which augment both cellular and humoral immune responses, and TH-2 cells that produce IL-4, IL-5 and IL-10 which augment humoral immune responses (Mosmann et al.,
J. Immunol.
126:2348 (1986)). It is often desirable to enhance the immunogenic potency of an antigen in order to obtain a stronger immune response in the organism being immunized and to strengthen host resistance to the antigen-bearing agent. A substance that enhances the immunogenicity of an antigen with which it is administered is known as an adjuvant. For example, certain lymphokines have been shown to have adjuvant activity, thereby enhancing the immune response to an antigen (Nencioni et al.,
J. Immunol.
139:800-804 (1987); EP285441 to Howard et al.).
SUMMARY OF THE INVENTION
This invention pertains to vaccine compositions comprising a mixture of herpes simplex virus glycoprotein D, the interleukin IL-12 and a mineral in suspension. The IL-12 may be either adsorbed onto the mineral suspension or simply mixed therewith. In a particular embodiment of the invention, the IL-12 is adsorbed onto a mineral suspension such as alum (e.g., aluminum hydroxide or aluminum phosphate). In a particular embodiment, the IL-12 is human IL-12. The invention also pertains to vaccine compositions which further comprise a physiologically acceptable vehicle. The invention further relates to immunogenic compositions comprising a mixture of a herpes simplex virus glycoprotein D, an adjuvant amount of interleukin-12, a mineral in suspension, and optionally comprising a physiologically acceptable vehicle.
The compositions of the present invention modulate the protective immune response to the antigen; that is, the vaccine composition is capable of quantitatively and qualitatively improving the vaccinated host's antibody response, and quantitatively increasing cell-mediated immunity for a protective response to a pathogen. In a particular embodiment of the invention, the antigen is a herpes simplex viral (HSV) antigen, such as envelope glycoprotein D (gD) of herpes simplex virus types I and/or II.
The invention also pertains to methods for preparing a vaccine composition comprising mixing HSV gD and IL-12 with a mineral in suspension. In particular, the IL-12 is adsorbed onto the mineral suspension. The invention also pertains to methods for eliciting or increasing a vaccinate's humoral and/or cell-mediated immunity, for a protective immune response, comprising administering to a vertebrate host an effective amount of a vaccine composition comprising a mixture of HSV gD, IL-12 and a mineral in suspension in a physiologically acceptable solution. In particular, the IL-12 is adsorbed onto the mineral suspension.
DETAILED DESCRIPTION OF THE INVENTION
Glycoprotein D (gD) is an envelope glycoprotein of herpes simplex virus (HSV) types I and II. HSV gD has been shown to be a potent inducer of protective immunity against primary and recurrent HSV infection in animal models (Mishkin et al.,
Vaccine
9:147-153 (1991); Landolfi et al.,
Vaccine
11:407-414 (1993)).
IL-12 is produced by a variety of antigen-presenting cells, principally macrophages and monocytes. It is a critical element in the induction of TH-1 cells from naive T cells. Production of IL-12 or the ability to respond to it has been shown to be critical in the development of protective TH-1-like responses, for example, during parasitic infections, most notably Leishmaniasis (Scott et al., U.S. Pat. No. 5,571,515). The effects of IL-12 are mediated by IFN-&ggr; produced by NK cells and T helper cells. IFN-&ggr; is critical for the induction of IgG2a antibodies to T-dependent protein antigens (Finkelman and Holmes,
Annu. Rev. Immunol.
8:303-33 (1990)) and IgG3 responses to T-independent antigens (Snapper et al.,
J. Exp. Med.
175:1367-1371 (1992)). Interleukin-12 (IL-12), originally called natural killer cell stimulatory factor, is a heterodimeric cytokine (Kobayashi et al.,
J. Exp. Med.
170:827 (1989)). The expression and isolation of IL-12 protein in recombinant host cells is described in International Patent Application WO 90/05147, published May 17, 1990.
The studies described herein relate to the utility of IL-12 as an adjuvant in a herpes simplex virus (HSV) vaccine. Accordingly, this invention pertains to vaccine compositions comprising a mixture of HSV gD, IL-12 and a mineral in suspension. In a particular embodiment of the invention, the IL-12 is adsorbed onto a mineral suspension such as alum (e.g., aluminum hydroxide or aluminum phosphate). These vaccine compositions modulate the protective immune response to HSV; that is, the vaccine composition is capable of eliciting the vaccinated host's cell-mediated immunity for a protective response to the pathogenic antigen.
IL-12 can be obtained from several suitable sources. It can be produced by recombinant DNA methodology; for example, the gene encoding human IL-12 has been cloned and expressed in host systems, permitting the production of large quantities of pure human IL-12. Also useful in the present invention are biologically active subunits or fragments of IL-12. Further, certain T lymphocyte lines produce high levels of IL-12, thus providing a readily available source. Commercial sources of recombinant human and murine IL-12 include Genetics Institute, Inc. (Cambridge, Mass.).
The antigen of this invention, e.g., an HSV antigen, can be used to elicit an immune response to she antigen in a vertebrate such as a mammalian host. For example, the antigen can be an HSV gD protein antigen or a portion thereof which retains the ability to stimulate an immune response.
The method of the present invention comprises administering to a mammal, particularly a human or other primate, an immunologically effective dose of a vaccine composition comprising a mixture of an antigen, e.g., an HSV gD antigen, an adjuvant amount of IL-12 and a mineral in suspension. In particular, the IL-12 is adsorbed onto the mineral suspension. As used herein, an “adjuvant amount” of IL-12 is intended to mean doses of from about 1 nanogram to about 20 micrograms, and more particularly from about 100 nanograms to about 5 micrograms. As used herein, an “immunologically effective” dose of the vaccine composition is a dose which is suitable to elicit an immune response. The particular dosage will depend upon the age, weight and medical condition of the mammal to be treated, as well as on the method of administration. Suitable doses will be readily determined by the skilled artisan. The vaccine composition can be optionally administered in a pharmaceutically or physiologically acceptable vehicle, such as physiological or phosphate buffered saline or ethanol polyols such as glycerol or propylene glycol. A small amount of detergent may also be included to enhance vaccine stability.
The vaccine composition may optionally comprise additional adjuvants such as vegetable oils or emulsions thereof, surface active substances, e.g., hexadecylamin, octadecyl amino acid esters, octadecylamine, lysolecithin, dimethyl-dioctadecylammonium bromide, N,N-dicoctadecyl-N′-N′bis(2-hydroxyethyl-propane diamine), methoxyhexadecylglycerol, and pluronic polyols; polyamines, e.g., pyran, dextransulfate, poly IC, carbopol; peptides, e g., muramly. dipeptide, dimethylglycine,

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