Methods and compositions for emergency post-infection treatment of anthrax

Abstract

Compositions effective for treating or preventing an anthrax infection in a subject in need thereof and recombinant proteins included in the compositions are provided. Methods for producing recombinant proteins in plants are described. Transgenic plants engineered to produce recombinant proteins as well as genetic constructs comprising nucleic acids encoding recombinant proteins thereof are also described. Methods of protecting subjects against anthrax infection with plant-derived compositions are provided.

1 . A genetic construct comprising a first polynucleotide and a second polynucleotide, wherein the first polynucleotide encodes a PA-LF protein, and the second polynucleotide encodes a carrier-protein selected from the group consisting of: Fc-IgA, Fc-IgG, Fc-IgM, oleosin, and VP2 coat protein. 
     
     
         2 . The genetic construct of  claim 8 , wherein the first polynucleotide includes a sequence with at least 90% identity to a reference sequence selected from the group consisting of: SEQ ID NO: 3 (PgA2-1), SEQ ID NO: 4(PgA2-2), SEQ ID NO: 5 (A2-3/PA-LF), and SEQ ID NO: 57 (PgA2-4). 
     
     
         3 . The genetic construct of  claim 8 , wherein the second polynucleotide includes a sequence with at least 90% identity to a reference sequence selected from the group consisting of: SEQ ID NO:6 (PgB1-1), SEQ ID NO: 7 (PgB2-1), SEQ ID NO: 8 (PgB3-1), SEQ ID NO: 9 (PgB1-2), SEQ ID NO: 10 (PgB2-2), SEQ ID NO: 11 (PgB3-2), SEQ ID NO: 12 (PgB4-1), SEQ ID NO: 13 (PgB5-1), SEQ ID NO: 48 (PgB1-3), SEQ ID NO: 50 (PgB1-4), SEQ ID NO: 51 (PgB2-3), SEQ ID NO: 52 (PgB2-4), SEQ ID NO: 53 (PgB3-3), SEQ ID NO: 54 (PgB3-4), SEQ ID NO: 55 (PgB4-2), and SEQ ID NO: 56 (PgB5-2). 
     
     
         4 . The genetic construct of  claim 1  further comprising a third polynucleotide encoding a helper element comprising a sequence with at least 90% identity to a reference sequence selected from the group consisting of: SEQ ID NO: 14 (PgC1-1), SEQ ID NO: 15 (PgC2-1), SEQ ID NO: 32 (PgC1-2), SEQ ID NO: 33 (PgC1-3), and SEQ ID NO: 34 (PgC2-2). 
     
     
         5 . The genetic construct of  claim 1  comprising a sequence with at least 90% identity to a reference sequence selected from the group consisting of: SEQ ID NO: 37 (PgA2-4:B1-5). 
     
     
         6 . A recombinant protein produced by the genetic construct of  claim 1 . 
     
     
         7 . A transgenic plant produced by the genetic construct of  claim 1 . 
     
     
         8 . The transgenic plant of  claim 7 , wherein the plant is a crop plant selected from the group consisting of: tomato, tobacco, pepper, eggplant, lettuce, oilseed rape, broccoli, cauliflower, cabbage crops, cucumber, melon, pumpkin, squash, peanut, soybeans, corn, rice, barley, buckwheat, sugar cane, cotton, beans, cassava, potatoes, sweet potatoes, and okra. 
     
     
         9 . The transgenic plant of  claim 7 , wherein the plant is a medicinal plant selected from the group consisting of:  Caragana sinica, Codonopsis pilosulae, Hedyotis diffusa, Houttuynia cordata, Lonicera japonica, Morinda officinalis, Oenothera odorata, Kalanchoe pinnata, Echinacea angustifolia, Calendula officinalis , and  Arthemis nobilis.    
     
     
         10 . A method for producing a recombinant protein in a plant, wherein the method comprises:
 contacting a plant with a genetic construct of  claim 1 ;   obtaining a plant including the genetic construct and expressing a recombinant protein, wherein the recombinant protein is encoded by the genetic construct.   
     
     
         11 . The method of  claim 10 , wherein the method further comprises isolating and purifying the recombinant protein from the plant. 
     
     
         12 . A method for preparing a composition effective for treating or preventing an anthrax infection in a subject, wherein the method comprises providing a recombinant protein produced by the method of  claim 10 . 
     
     
         13 . The method of  claim 12 , wherein the method further comprises providing a pharmaceutically acceptable carrier. 
     
     
         14 . The method of  claim 12 , wherein the method further comprises providing an adjuvant. 
     
     
         15 . A method of protecting a subject against anthrax infection, wherein the method comprises: providing a composition that includes a recombinant protein of  claim 6 , wherein the composition is effective in preventing or reducing at least one symptom of an anthrax infection in a subject; and administering the composition to the subject in need thereof. 
     
     
         16 . The method of  claim 15 , wherein the subject is a mammal. 
     
     
         17 . The method of  claim 15 , wherein the mammal is selected from the group consisting of: an agricultural animal, an equine, a high value zoo animal, a research animal and a human. 
     
     
         18 . The method of  claim 15 , wherein administering includes a route selected from the group consisting of: intravenous, intramuscular, intraperitoneal, intradermal, mucosal, cutaneous and subcutaneous. 
     
     
         19 . The method of  claim 15 , wherein administering is intranasal. 
     
     
         20 . The method of  claim 19 , wherein intranasal administering is provided in the form of inhalation or nasal drops.