HCoV VACCINE FOR IMPROVING IMMUNITY AGAINST SARS-COV-2 INFECTION

Abstract

Embodiments include a method of using inactivated human cold coronaviruses (HCoVs) particularly HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1, alone or as a booster, for the immunization against SARS-CoV-2 infections. Vaccine embodiments further comprise HCoV virus envelope subunits which may be in the form of virus-like spheroids (VLS).

1 . A method for providing a pharmaceutical immunogenic composition comprising the steps of: (a) providing a plurality of population of cells in cell culture medium; (b) infecting each population of cells by inoculating the population of cells with the at least one of: HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 and incubating the inoculated population of cells to allow the virus in each cell culture medium to replicate and propagate; (c) collecting the virus from each cell culture medium; (d) purifying each of said virus from each cell culture; (e) inactivating each of said virus by at least one of beta-propiolactone (BPL), hydrogen peroxide, formalin (formaldehyde) and copper; and (f) preparing a pharmaceutical preparation for inoculation comprising viruses consisting of two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 inactivated whole virus of steps (a)-(e). 
     
     
         2 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-229E and HCoV-OC43. 
     
     
         3 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-229E and HCoV-NL63. 
     
     
         4 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-229E and HCoV-HKU1. 
     
     
         5 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-0C43 and HCoV-NL63. 
     
     
         6 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-OC43 and HCoV-HKU1. 
     
     
         7 . The method of  claim 1  wherein the two of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-NL63 and HCoV-HKU1. 
     
     
         8 . The method of  claim 1  wherein the cells infected at step (b) are Vero cells. 
     
     
         9 . The method of  claim 1  further wherein the pharmaceutical composition prepared at step (f) comprises an adjuvant from one at least of alum, algammulin, monophosphoryl lipid A (MPL), resiquimod, muramyl depeptide (MPD), N glycolyl dipeptide (GMDP), polylC, CpG oligonucleotide, aluminum salts, water in oil emulsion and oil in water emulsion. 
     
     
         10 . A method for providing a pharmaceutical immunogenic composition comprising the steps of: (a) providing a plurality of population of cells in cell culture medium; (b) infecting each population of cells by inoculating the population of cells with the at least one of: HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 and incubating the inoculated population of cells to allow the virus in each cell culture medium to replicate and propagate; (c) collecting the virus from each cell culture medium; (d) purifying each of said virus from each cell culture; (e) inactivating each of said virus by at least one of beta-propiolactone (BPL), hydrogen peroxide, formalin (formaldehyde) and copper; and (f) preparing a pharmaceutical preparation for inoculation comprising viruses consisting of three of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 inactivated whole virus of steps (a)-(e). 
     
     
         11 . The method of  claim 10  wherein the pharmaceutical composition prepared at step (f) further comprises an adjuvant from at least one of : alum, algammulin, monophosphoryl lipid A (MPL), resiquimod, muramyl depeptide (MPD), N glycolyl dipeptide (GMDP), polylC, CpG oligonucleotide, aluminum salts, water in oil emulsion and oil in water emulsion. 
     
     
         12 . The method of  claim 10  wherein the cells infected at step (b) are Vero cells. 
     
     
         13 . The method of  claim 10  wherein the three of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-229E, HCoV-NL63 and HCoV-HKU1. 
     
     
         14 . The method of  claim 9  wherein the three HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-OC43, HCoV-NL63 and HCoV-HKU1. 
     
     
         15 . The method of  claim 9  wherein the three of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-229E, HCoV-OC43, and HCoV-HKU1. 
     
     
         16 . The method of  claim 9  wherein the three of HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 consists of HCoV-NL63, HCoV-229 E, and HCoV-OC43. 
     
     
         17 . A method for providing a pharmaceutical immunogenic composition comprising the steps of: (a) providing a plurality of population of cells in cell culture medium; (b) infecting each population of cells by inoculating the population of cells with the at least one of: HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 and incubating the inoculated population of cells to allow the virus in each cell culture medium to replicate and propagate; (c) collecting the virus from each cell culture medium; (d) purifying each of said virus from each cell culture; (e) inactivating each of said virus by at least one of beta-propiolactone (BPL), hydrogen peroxide, formalin (formaldehyde) and copper; and (f) preparing a pharmaceutical preparation for inoculation comprising viruses consisting of all of—HCoV-299E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 inactivated whole virus of steps (a)-(e). 
     
     
         18 . The method of  claim 17  wherein the pharmaceutical composition prepared at step (f) further comprises an adjuvant from at least one of : alum, algammulin, monophosphoryl lipid A (MPL), resiquimod, muramyl depeptide (MPD), N glycolyl dipeptide (GMDP), polylC, CpG oligonucleotide, aluminum salts, water in oil emulsion and oil in water emulsion. 
     
     
         19 . The method of  claim 17  wherein the cells infected at step (b) are Vero cells.