Patents by Inventor Manuel V. Borca

Manuel V. Borca has filed for patents to protect the following inventions. This listing includes patent applications that are pending as well as patents that have already been granted by the United States Patent and Trademark Office (USPTO).

  • Publication number: 20210244809
    Abstract: Provided herein are details on the construction of a recombinant African Swine Fever Virus (ASFV) live attenuated vaccine for prevention of ASF caused by various strains of ASFV, such as the highly virulent Georgia 2007 isolate (“ASFV-G”). An exemplary vaccine comprises the ASFV-G?I1771 modified virus, a recombinant ASFV-G modified by deleting a portion of the I177L ORF rendering the I177L gene nonfunctional.
    Type: Application
    Filed: April 6, 2021
    Publication date: August 12, 2021
    Inventors: Douglas P. GLADUE, MANUEL V. BORCA
  • Patent number: 11007263
    Abstract: Provided herein are details on the construction of a recombinant African Swine Fever Virus (ASFV) live attenuated vaccine for prevention of ASF caused by various strains of ASFV, such as the highly virulent Georgia 2007 isolate (“ASFV-G”). An exemplary vaccine comprises the ASFV-G?I1771 modified virus, a recombinant ASFV-G modified by deleting a portion of the I177L ORF rendering the I177L gene nonfunctional.
    Type: Grant
    Filed: September 24, 2019
    Date of Patent: May 18, 2021
    Assignee: The United States of America, as represented by The Secretary of Agriculture
    Inventors: Douglas P. Gladue, Manuel V. Borca
  • Publication number: 20210085776
    Abstract: Provided herein are details on the construction of a recombinant African Swine Fever Virus (ASFV) live attenuated vaccine for prevention of ASF caused by various strains of ASFV, such as the highly virulent Georgia 2007 isolate (“ASFV-G”). An exemplary vaccine comprises the ASFV-G?I1771 modified virus, a recombinant ASFV-G modified by deleting a portion of the I177L ORF rendering the I177L gene nonfunctional.
    Type: Application
    Filed: September 24, 2019
    Publication date: March 25, 2021
    Inventors: Douglas P. GLADUE, MANUEL V. BORCA
  • Patent number: 9814771
    Abstract: The role of a specific E2 region containing a putative fusion peptide (FP) sequence was evaluated. FPs critically contribute to the interaction between proteins and the membrane system of the host cell. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how amino acid substitutions within this region of E2 may affect replication of BICv in cell cultures and affect virus virulence in swine. Interestingly, mutated virus FPi.c was completely attenuated when inoculated intranasally at a dose of 105 TCID50 in swine. Importantly, animals infected with FPi.c virus were protected against the virulent challenge with Brescia virus at 3 and 28 days after vaccination. Protection was evidenced by absence of clinical signs related with CSF as well as the absence of viremia produced by the challenge virulent virus.
    Type: Grant
    Filed: September 8, 2016
    Date of Patent: November 14, 2017
    Assignees: The United States of America, as represented by The Secretary of Agriculture, The University of Connecticut, Universidad del Pais Vasco/Euskal Herriko Univertsitatea (UPV-EHU)
    Inventors: Manuel V. Borca, Douglas P. Gladue, Lauren G. Holinka-Patterson, Vivian O'Donnell, Jose Nieva
  • Patent number: 9808520
    Abstract: African swine fever virus (ASFV) is the etiological agent of a contagious, often lethal viral disease of domestic pigs. The control of African Swine Fever (ASF) has been hampered by the unavailability of vaccines. Experimental vaccines have been derived from naturally occurring, cell culture-adapted, or genetically modified live attenuated ASFVs; however, these vaccines are only successful when protecting against homologous viruses. We have constructed a recombinant ?9GL/?UK virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate by deleting the specific virulence-associated 9GL (B119L) and the UK (DP96R) genes. In vivo, ASFV-G ?9GL/?UK administered intramuscularly to swine even at relatively high doses (106 HAD50) does not induce disease. Importantly, animals infected with 104 or 106 HAD50 are solidly protected against the presentation of clinical disease when challenged at 28 days post infection with the virulent parental strain Georgia 2007.
    Type: Grant
    Filed: July 1, 2016
    Date of Patent: November 7, 2017
    Assignees: The United States of America as represented by The Secretary of Agriculture, The University of Connecticut
    Inventors: Manuel V. Borca, Douglas P. Gladue, Lauren G. Holinka-Patterson, Guillermo R. Risatti, Vivian K. O'Donnell
  • Publication number: 20170128563
    Abstract: The role of a specific E2 region, 869CKWGGNWTCV878, containing a putative fusion peptide (FP) sequence was evaluated. FPs critically contribute to the interaction between proteins and the membrane system of the host cell. Reverse genetics utilizing a full-length infectious clone of the highly virulent CSFV strain Brescia (BICv) was used to evaluate how amino acid substitutions within this region of E2 may affect replication of BICv in cell cultures and affect virus virulence in swine. A recombinant CSFV (FPi.c) containing mutations in three amino acid residues within the E2 protein area comprised CSFV amino acid residues 869-878 was constructed: W871T, W875D, and V878T. Interestingly, mutated virus FPi.c was completely attenuated when inoculated intranasally at a dose of 105 TCID50 in swine. Importantly, animals infected with FPi.c virus were protected against the virulent challenge with Brescia virus at 3 and 28 days after vaccination.
    Type: Application
    Filed: September 8, 2016
    Publication date: May 11, 2017
    Inventors: Manuel V. Borca, Douglas P. Gladue, Lauren G. Holinka-Patterson, Vivian O'Donnell, Jose Nieva
  • Patent number: 9528094
    Abstract: African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs. Control of ASF has been hampered by the unavailability of vaccines. Experimental vaccines have been derived from naturally occurring, cell culture-adapted, or genetically modified live attenuated ASFVs; however, these vaccines are only successful when protecting against homologous viruses. Among viral genes reported to be involved in virulence are components of the multi gene family (MGF). Here we report the construction of a recombinant ?MGF virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate. In vivo, ASFV-G ?MGF administered intramuscularly (IM) to swine at either 102 or 104 HAD50 are completely attenuated; the inoculated animals are completely asymptomatic. Animals infected with 102 or 104 HAD50 of ASFV-G ?MGF are protected against the presentation of clinical disease when challenged at 28 days post infection with the virulent parental strain Georgia 2007.
    Type: Grant
    Filed: November 10, 2014
    Date of Patent: December 27, 2016
    Assignees: The United States of America, as Represented by the Secretary of Agriculture, The University of Connecticut
    Inventors: Manuel V. Borca, Lauren G. Holinka-Patterson, Vivian K. O'Donnell, Guillermo S. Risatti, Douglas Gladu
  • Patent number: 9474797
    Abstract: We have developed an ASFV Georgia strain adapted to grow in Vero cell line. The resulting virus, ASF-GVAV, efficiently grows in Vero cells although it still is able to significantly replicate in primary cell cultures of swine macrophages. ASF-GVAV virus was successfully used as parental virus to develop several recombinant ASF viruses. The development of an ASFV adapted to grow in an established cell line is a significant advance for research and development of vaccine candidate strains using genetic manipulation based in the process of homologous recombination. The GVAVS can be utilized as a basis for large scale production of ASF vaccines.
    Type: Grant
    Filed: June 19, 2014
    Date of Patent: October 25, 2016
    Assignees: The United States of America, as Represented by the Secretary of Agriculture, The University of Connecticut
    Inventors: Manuel V. Borca, Luis L. Rodriguez, Peter W. Krug, Vivian K. O'Donnell
  • Patent number: 9463234
    Abstract: African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs that has significant economic consequences for swine breeding. Control of ASF has been hampered by the unavailability of vaccines. Recombinant viruses harboring engineered deletions of specific virulence-associated genes induce solid protection against the challenge with parental viruses. Here we report the construction of a recombinant ?9GL virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate. In vivo, ASFV-G ?9GL administered intramuscularly (IM) to swine at relatively high doses (104 HAD50) retains a virulent phenotype practically indistinguishable from the parental virus. Conversely, at low IM doses (102 or 103 HAD50), ASFV-G ?9GL does not induce disease. Importantly, animals infected with 103 HAD50 are protected against the presentation of clinical disease when challenge at 28 days post infection with the virulent parental strain Georgia 2007.
    Type: Grant
    Filed: September 24, 2014
    Date of Patent: October 11, 2016
    Assignees: The United States of America, as represented by the Secretary of Agriculture, University of Connecticut
    Inventors: Manuel V. Borca, Lauren G. Holinka-Patterson, Douglas Gladue, Guillermo R. Risatti, Vivian K. O'Donnell
  • Patent number: 9352032
    Abstract: Controlling Classical Swine Fever Virus (CSFV) involves either prophylactic vaccination or non-vaccination and elimination of infected herds depending on the epidemiological situation. Marker vaccines allowing distinction between naturally infected from vaccinated swine could complement “stamping out” measures. Previously, we reported the development of FlagT4v, a double antigenic marker live attenuated CSFV strain. FlagT4v was later shown as not to be completely stable in terms of its attenuation when assessed in a reversion to virulence protocol. We have developed a modified version of the FlagT4v where changes in the codon usage of genomic areas encoding for Flag and T4 were introduced to rectify the reversion to the virulent genotype. The new virus, FlagT4-mFT-Gv, possesses the same amino acid sequence as FlagT4v except for one substitution, Asparagine is replaced by Glycine at position 852 of the CSFV polypeptide.
    Type: Grant
    Filed: March 7, 2014
    Date of Patent: May 31, 2016
    Assignees: The United States of America as represented by The Secretary of Agriculture, The University of Conneticut
    Inventors: Manuel V. Borca, Guillermo R. Risatti, Lauren G. Holinka-Patterson
  • Publication number: 20160130562
    Abstract: African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs. Control of ASF has been hampered by the unavailability of vaccines. Experimental vaccines have been derived from naturally occurring, cell culture-adapted, or genetically modified live attenuated ASFVs; however, these vaccines are only successful when protecting against homologous viruses. Among viral genes reported to be involved in virulence are components of the multi gene family (MGF). Here we report the construction of a recombinant ?MGF virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate. In vivo, ASFV-G ?MGF administered intramuscularly (IM) to swine at either 102 or 104 HAD50 are completely attenuated; the inoculated animals are completely asymptomatic. Animals infected with 102 or 104 HAD50 of ASFV-G ?MGF are protected against the presentation of clinical disease when challenged at 28 days post infection with the virulent parental strain Georgia 2007.
    Type: Application
    Filed: November 10, 2014
    Publication date: May 12, 2016
    Inventors: Manuel V. Borca, Lauren G. Holinka-Patterson, Vivian K. O'Donnell, Guillermo S. Risatti, Douglas Gladu
  • Publication number: 20160082099
    Abstract: African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal viral disease of domestic pigs that has significant economic consequences for swine breeding. Control of ASF has been hampered by the unavailability of vaccines. Recombinant viruses harboring engineered deletions of specific virulence-associated genes induce solid protection against the challenge with parental viruses. Here we report the construction of a recombinant ?9GL virus derived from the highly virulent ASFV Georgia 2007 (ASFV-G) isolate. In vivo, ASFV-G ?9GL administered intramuscularly (IM) to swine at relatively high doses (104 HAD50) retains a virulent phenotype practically indistinguishable from the parental virus. Conversely, at low IM doses (102 or 103 HAD50), ASFV-G ?9GL does not induce disease. Importantly, animals infected with 103 HAD50 are protected against the presentation of clinical disease when challenge at 28 days post infection with the virulent parental strain Georgia 2007.
    Type: Application
    Filed: September 24, 2014
    Publication date: March 24, 2016
    Inventors: Manuel V. Borca, Lauren G. Holinka-Patterson, Douglas Gladue, Guillermo R. Risatti, Vivian K. O'Donnell
  • Publication number: 20150250866
    Abstract: Controlling Classical Swine Fever Virus (CSFV) involves either prophylactic vaccination or non-vaccination and elimination of infected herds depending on the epidemiological situation. Marker vaccines allowing distinction between naturally infected from vaccinated swine could complement “stamping out” measures. Previously, we reported the development of FlagT4v, a double antigenic marker live attenuated CSFV strain. FlagT4v was later shown as not to be completely stable in terms of its attenuation when assessed in a reversion to virulence protocol. We have developed a modified version of the FlagT4v where changes in the codon usage of genomic areas encoding for Flag and T4 were introduced to rectify the reversion to the virulent genotype. The new virus, FlagT4-mFT-Gv, possesses the same amino acid sequence as FlagT4v except for one substitution, Asparagine is replaced by Glycine at position 852 of the CSFV polypeptide.
    Type: Application
    Filed: March 7, 2014
    Publication date: September 10, 2015
    Inventors: Manuel V. Borca, Guillermo R. Risatti, Lauren G. Holinka-Patterson
  • Patent number: 8846055
    Abstract: Classical Swine Fever Virus (CSFV) E2 glycoprotein is a major inducer of neutralizing antibodies and protective immunity in swine. E2 mediates virus adsorption to the target cell, and harbors genetic determinants associated with virus virulence. CSFV E2 also contains between residues 829 and 837 a discrete epitope (TAVSPTTLR) recognized by monoclonal antibody (mAb) WH303, used to differentiate CSFV from related Pestiviruses Bovine Viral Diarrhea Virus (BVDV) and Border Disease Virus (BDV). In this report, a CSFV infectious clone of the virulent Brescia isolate (BICv) was used to progressively mutate the mAb WH303 epitope of CSFV E2 to the homologous amino acid sequence of BVDV strain NADL E2 (TSFNMDTLA).
    Type: Grant
    Filed: May 30, 2006
    Date of Patent: September 30, 2014
    Assignee: The United States of America, as Represented by the Secretary of Agriculture
    Inventors: Manuel V. Borca, Guillermo R. Risatti
  • Patent number: 8426575
    Abstract: E1, along with Erns and E2 is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). Our previous studies indicated that glycosylation status of either E2 or Erns strongly influence viral virulence in swine. Here, we have investigated the role of E1 glycosylation of highly virulent CSFV strain Brescia during infection in the natural host. The three putative glycosylation sites in E1 were modified by site directed mutagenesis of a CSFV Brescia infectious clone (BICv). A panel of virus mutants was obtained and used to investigate whether the removal of putative glycosylation sites in the E1 glycoprotein would affect viral virulence/pathogenesis in swine. We observed that rescue of viable virus was completely impaired by removal of all three putative glycosylation sites in E1. Single mutations of each of the E1 glycosylation sites showed that CSFV amino acid N594 (E1.N3 virus), as well the combined mutation of N500 and N513 (E1.N1N2 virus) resulted in BICv attenuation.
    Type: Grant
    Filed: September 21, 2011
    Date of Patent: April 23, 2013
    Assignee: The United States of America, as Represented by the Secretary of Agriculture
    Inventors: Manuel V. Borca, Guillermo R. Risatti
  • Patent number: 8133495
    Abstract: Classical swine fever virus is a world-wide distributed highly-contagious disease affecting swine. The two main strategies for diseases control are prophylactic vaccination and non-vaccination stamping out policies. Marker vaccines are a promising strategy. Here we report the rational development of a doubly antigenic marker CSFV experimental live attenuated candidate strain vaccine (Flag/T4 virus). Flag/T virus (Flag/T4v) is based in the combination of two Brescia derived recombinant attenuated viruses: RB-C22 and T4. RB-C22v contains a 19mer insertion in the structural glycoprotein E1, while T4v posses mutated CSFV amino acid residues 830 to 834 in the structural glycoprotein E2, deleting the highly conserved epitope recognized by monoclonal antibody (mAb) WH303. Flag/T4 virus contains a positive foreign antigenic marker, due to the insertion of the highly antigenic epitope Flag in the 19mer insertion of E1, as well as a negative antigenic marker, the lack of reactivity with mAb WH303.
    Type: Grant
    Filed: May 22, 2007
    Date of Patent: March 13, 2012
    Assignees: The United States of America as represented by the Secretary of Agriculture, The University of Connecticut
    Inventors: Manuel V. Borca, Guillermo R. Risatti
  • Patent number: 8063195
    Abstract: NS4B is one of the non-structural proteins of classical swine fever virus. By using functional genetics, we have discovered, in the predicted amino acid sequence of NS4B of CSFV strain Brescia, a motif that resembles those found in the toll-like receptor (TLR) proteins, a group of host cell proteins involved in the development of anti-viral mechanisms. We have located the TLR motif in two groups of amino acid triplets at amino acid positions 2531-3 (residues IYK) and 2566-8 (residues VGI) of the CSFV NS4B glycoprotein. We have constructed a recombinant CSFV (derived from an infectious clone containing the genetic information of the highly virulent strain Brescia) containing amino acid substitutions in the three amino acid residues at positions 2566, 2567 and 2568, where the VGI triplet has been replaced by an AAA triplet inside the NS4B glycoprotein. The obtained virus, named NS4B-VGIv, was completely attenuated in swine, showing a limited ability in spreading during the infection in vivo.
    Type: Grant
    Filed: May 22, 2009
    Date of Patent: November 22, 2011
    Assignee: The United States of America as represented by the Secretary of Agriculture
    Inventors: Manuel V. Borca, James J. Zhu
  • Publication number: 20100297175
    Abstract: NS4B is one of the non-structural proteins of classical swine fever virus. By using functional genetics, we have discovered, in the predicted amino acid sequence of NS4B of CSFV strain Brescia, a motif that resembles those found in the toll-like receptor (TLR) proteins, a group of host cell proteins involved in the development of anti-viral mechanisms. We have located the TLR motif in two groups of amino acid triplets at amino acid positions 2531-3 (residues IYK) and 2566-8 (residues VGI) of the CSFV NS4B glycoprotein. We have constructed a recombinant CSFV (derived from an infectious clone containing the genetic information of the highly virulent strain Brescia) containing amino acid substitutions in the three amino acid residues at positions 2566, 2567 and 2568, where the VGI triplet has been replaced by an AAA triplet inside the NS4B glycoprotein. The obtained virus, named NS4B-VGIv, was completely attenuated in swine, showing a limited ability in spreading during the infection in vivo.
    Type: Application
    Filed: May 22, 2009
    Publication date: November 25, 2010
    Inventors: Manuel V. Borca, James J. Zhu
  • Publication number: 20080292653
    Abstract: Classical swine fever virus is a world-wide distributed highly-contagious disease affecting swine. The two main strategies for diseases control are prophylactic vaccination and non-vaccination stamping out policies. Marker vaccines are a promising strategy. Here we report the rational development of a doubly antigenic marker CSFV experimental live attenuated candidate strain vaccine (Flag/T4 virus). Flag/T virus (Flag/T4v) is based in the combination of two Brescia derived recombinant attenuated viruses: RB-C22 and T4. RB-C22v contains a 19mer insertion in the structural glycoprotein E1, while T4v posses mutated CSFV amino acid residues 830 to 834 in the structural glycoprotein E2, deleting the highly conserved epitope recognized by monoclonal antibody (mAb) WH303. Flag/T4 virus contains a positive foreign antigenic marker, due to the insertion of the highly antigenic epitope Flag in the 19mer insertion of E1, as well as a negative antigenic marker, the lack of reactivity with mAb WH303.
    Type: Application
    Filed: May 22, 2007
    Publication date: November 27, 2008
    Inventors: Manuel V. Borca, Guillermo R. Risatti
  • Patent number: 7332170
    Abstract: Transposon linker insertion mutagenesis of a full-length infectious clone of the highly pathogenic classical swine fever virus (CSFV) isolate Brescia (pBIC) was used to identify genetic determinants of CSFV virulence and host range. A virus mutant, RB-C22 (RB-C22v), possessing a 19-residue tag insertion at the carboxyl end of E1 was constructed. RB-C22v and the parental virus pBIC (pBICv) exhibited similar growth characteristics on primary porcine macrophage cell cultures although RB-C22v produced significantly smaller plaques on SK6 cell cultures. In vivo, RB-C22v was markedly attenuated in swine. In contrast with pBIC infection, where mortality was 100%, all RB-C22v-infected pigs survived infection remaining clinically normal. Additionally, chimeras of the Brescia strain and the attenuated vaccine strain CS were constructed and evaluated for viral virulence in swine. Chimeras 138.8v and 337.14v, chimeras containing the E2 glycoprotein of CS and chimeric virus 319.
    Type: Grant
    Filed: December 23, 2005
    Date of Patent: February 19, 2008
    Assignee: The United States of America, as represented by the Secretary of Agriculture
    Inventors: Manuel V. Borca, Guillermo R. Risatti, Daniel L. Rock