WO1999038520A9 - Blood cell composition as a vaccine for hiv and other pathogenic organisms - Google Patents
Blood cell composition as a vaccine for hiv and other pathogenic organismsInfo
- Publication number
- WO1999038520A9 WO1999038520A9 PCT/US1999/001922 US9901922W WO9938520A9 WO 1999038520 A9 WO1999038520 A9 WO 1999038520A9 US 9901922 W US9901922 W US 9901922W WO 9938520 A9 WO9938520 A9 WO 9938520A9
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- composition
- cells
- blood
- hiv
- viruses
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 177
- 229960005486 vaccine Drugs 0.000 title claims abstract description 9
- 210000000601 blood cell Anatomy 0.000 title claims description 7
- 244000052769 pathogen Species 0.000 title description 2
- 210000004369 blood Anatomy 0.000 claims abstract description 65
- 239000008280 blood Substances 0.000 claims abstract description 65
- 238000000034 method Methods 0.000 claims abstract description 52
- 241000282414 Homo sapiens Species 0.000 claims abstract description 46
- 238000011282 treatment Methods 0.000 claims abstract description 26
- 238000002360 preparation method Methods 0.000 claims abstract description 25
- 239000003814 drug Substances 0.000 claims abstract description 24
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 16
- 201000010099 disease Diseases 0.000 claims abstract description 14
- 230000000840 anti-viral effect Effects 0.000 claims abstract description 12
- 231100000252 nontoxic Toxicity 0.000 claims abstract description 5
- 230000003000 nontoxic effect Effects 0.000 claims abstract description 5
- 230000002519 immonomodulatory effect Effects 0.000 claims abstract description 3
- 230000001613 neoplastic effect Effects 0.000 claims abstract 2
- 229940124597 therapeutic agent Drugs 0.000 claims abstract 2
- 210000004027 cell Anatomy 0.000 claims description 144
- 241001504519 Papio ursinus Species 0.000 claims description 50
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 claims description 49
- 241000700605 Viruses Species 0.000 claims description 45
- 230000000694 effects Effects 0.000 claims description 44
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 43
- 241000725303 Human immunodeficiency virus Species 0.000 claims description 28
- 210000000265 leukocyte Anatomy 0.000 claims description 28
- 235000002639 sodium chloride Nutrition 0.000 claims description 25
- 239000000243 solution Substances 0.000 claims description 25
- 239000011780 sodium chloride Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 230000003612 virological effect Effects 0.000 claims description 20
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 19
- 102000004169 proteins and genes Human genes 0.000 claims description 19
- 108090000623 proteins and genes Proteins 0.000 claims description 19
- 210000003743 erythrocyte Anatomy 0.000 claims description 18
- 210000001519 tissue Anatomy 0.000 claims description 18
- 241001465754 Metazoa Species 0.000 claims description 14
- 241000282520 Papio Species 0.000 claims description 13
- 208000015181 infectious disease Diseases 0.000 claims description 13
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 12
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 12
- 241000288906 Primates Species 0.000 claims description 11
- 230000000087 stabilizing effect Effects 0.000 claims description 11
- 210000000130 stem cell Anatomy 0.000 claims description 10
- 230000002779 inactivation Effects 0.000 claims description 9
- 150000001875 compounds Chemical class 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 210000004700 fetal blood Anatomy 0.000 claims description 8
- 210000004698 lymphocyte Anatomy 0.000 claims description 8
- 241000894007 species Species 0.000 claims description 8
- 230000001225 therapeutic effect Effects 0.000 claims description 7
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 6
- 229930006000 Sucrose Natural products 0.000 claims description 6
- 239000001110 calcium chloride Substances 0.000 claims description 6
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
- 239000001103 potassium chloride Substances 0.000 claims description 6
- 235000011164 potassium chloride Nutrition 0.000 claims description 6
- 229910000160 potassium phosphate Inorganic materials 0.000 claims description 6
- 235000011009 potassium phosphates Nutrition 0.000 claims description 6
- 239000001488 sodium phosphate Substances 0.000 claims description 6
- 229910000162 sodium phosphate Inorganic materials 0.000 claims description 6
- 235000011008 sodium phosphates Nutrition 0.000 claims description 6
- 239000005720 sucrose Substances 0.000 claims description 6
- 239000003104 tissue culture media Substances 0.000 claims description 6
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 claims description 6
- 230000027455 binding Effects 0.000 claims description 5
- 235000011148 calcium chloride Nutrition 0.000 claims description 5
- 230000003833 cell viability Effects 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 230000001105 regulatory effect Effects 0.000 claims description 5
- 239000003146 anticoagulant agent Substances 0.000 claims description 4
- 229940127219 anticoagulant drug Drugs 0.000 claims description 4
- 229960004222 factor ix Drugs 0.000 claims description 4
- 210000001616 monocyte Anatomy 0.000 claims description 4
- 241001529453 unidentified herpesvirus Species 0.000 claims description 4
- 102100022641 Coagulation factor IX Human genes 0.000 claims description 3
- 241000450599 DNA viruses Species 0.000 claims description 3
- 108010076282 Factor IX Proteins 0.000 claims description 3
- 230000002411 adverse Effects 0.000 claims description 3
- 239000011575 calcium Substances 0.000 claims description 3
- 150000001720 carbohydrates Chemical class 0.000 claims description 3
- 235000014633 carbohydrates Nutrition 0.000 claims description 3
- 241001493065 dsRNA viruses Species 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 208000006454 hepatitis Diseases 0.000 claims description 3
- 231100000283 hepatitis Toxicity 0.000 claims description 3
- 239000000815 hypotonic solution Substances 0.000 claims description 3
- 244000005700 microbiome Species 0.000 claims description 3
- 244000309711 non-enveloped viruses Species 0.000 claims description 3
- 229920000642 polymer Polymers 0.000 claims description 3
- 230000005855 radiation Effects 0.000 claims description 3
- GTJOHISYCKPIMT-UHFFFAOYSA-N 2-methylundecane Chemical compound CCCCCCCCCC(C)C GTJOHISYCKPIMT-UHFFFAOYSA-N 0.000 claims description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 2
- -1 Factor VIIII Proteins 0.000 claims description 2
- 229910052791 calcium Inorganic materials 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- 230000000120 cytopathologic effect Effects 0.000 claims description 2
- 230000007812 deficiency Effects 0.000 claims description 2
- 230000001605 fetal effect Effects 0.000 claims description 2
- 210000005087 mononuclear cell Anatomy 0.000 claims description 2
- 210000000056 organ Anatomy 0.000 claims description 2
- 239000000137 peptide hydrolase inhibitor Substances 0.000 claims description 2
- 238000010257 thawing Methods 0.000 claims description 2
- 231100000676 disease causative agent Toxicity 0.000 claims 4
- 230000000415 inactivating effect Effects 0.000 claims 4
- 230000002934 lysing effect Effects 0.000 claims 4
- 210000001185 bone marrow Anatomy 0.000 claims 3
- 230000000845 anti-microbial effect Effects 0.000 claims 2
- 239000012503 blood component Substances 0.000 claims 2
- 238000002255 vaccination Methods 0.000 claims 2
- 206010027654 Allergic conditions Diseases 0.000 claims 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims 1
- 102100023804 Coagulation factor VII Human genes 0.000 claims 1
- 108010023321 Factor VII Proteins 0.000 claims 1
- 241000233866 Fungi Species 0.000 claims 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 206010061598 Immunodeficiency Diseases 0.000 claims 1
- 208000029462 Immunodeficiency disease Diseases 0.000 claims 1
- 229910019142 PO4 Inorganic materials 0.000 claims 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 claims 1
- 241000125945 Protoparvovirus Species 0.000 claims 1
- 239000004480 active ingredient Substances 0.000 claims 1
- 230000003266 anti-allergic effect Effects 0.000 claims 1
- 230000003110 anti-inflammatory effect Effects 0.000 claims 1
- 229960005475 antiinfective agent Drugs 0.000 claims 1
- 239000004599 antimicrobial Substances 0.000 claims 1
- 244000052616 bacterial pathogen Species 0.000 claims 1
- 210000000170 cell membrane Anatomy 0.000 claims 1
- 239000006071 cream Substances 0.000 claims 1
- 229940012413 factor vii Drugs 0.000 claims 1
- 244000000011 human parasite Species 0.000 claims 1
- 230000007813 immunodeficiency Effects 0.000 claims 1
- 230000004968 inflammatory condition Effects 0.000 claims 1
- 239000006210 lotion Substances 0.000 claims 1
- 238000001471 micro-filtration Methods 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 1
- 239000010452 phosphate Substances 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
- 230000003449 preventive effect Effects 0.000 claims 1
- 239000003419 rna directed dna polymerase inhibitor Substances 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 238000000527 sonication Methods 0.000 claims 1
- 235000000346 sugar Nutrition 0.000 claims 1
- 235000021092 sugar substitutes Nutrition 0.000 claims 1
- 150000008163 sugars Chemical class 0.000 claims 1
- 239000003765 sweetening agent Substances 0.000 claims 1
- 230000000699 topical effect Effects 0.000 claims 1
- 208000030507 AIDS Diseases 0.000 abstract description 15
- 102100034343 Integrase Human genes 0.000 abstract description 5
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 abstract description 5
- 230000002401 inhibitory effect Effects 0.000 abstract description 5
- 108091005804 Peptidases Proteins 0.000 abstract description 4
- 239000004365 Protease Substances 0.000 abstract description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 4
- 230000003389 potentiating effect Effects 0.000 abstract description 4
- 208000003669 immune deficiency disease Diseases 0.000 abstract description 2
- 230000000903 blocking effect Effects 0.000 abstract 1
- 230000001404 mediated effect Effects 0.000 abstract 1
- 239000006166 lysate Substances 0.000 description 45
- 238000010790 dilution Methods 0.000 description 38
- 239000012895 dilution Substances 0.000 description 38
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 23
- 238000004458 analytical method Methods 0.000 description 22
- 238000003556 assay Methods 0.000 description 20
- 229940079593 drug Drugs 0.000 description 20
- 239000002953 phosphate buffered saline Substances 0.000 description 20
- 239000000523 sample Substances 0.000 description 20
- 238000012360 testing method Methods 0.000 description 19
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 18
- 230000005764 inhibitory process Effects 0.000 description 18
- 239000000284 extract Substances 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 13
- 208000031886 HIV Infections Diseases 0.000 description 12
- 210000000988 bone and bone Anatomy 0.000 description 12
- 238000001228 spectrum Methods 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 10
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 9
- 239000000427 antigen Substances 0.000 description 9
- 108091007433 antigens Proteins 0.000 description 9
- 102000036639 antigens Human genes 0.000 description 9
- 238000004128 high performance liquid chromatography Methods 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 229910001868 water Inorganic materials 0.000 description 9
- 241000282412 Homo Species 0.000 description 8
- 150000001413 amino acids Chemical class 0.000 description 8
- 210000000987 immune system Anatomy 0.000 description 8
- 208000037357 HIV infectious disease Diseases 0.000 description 7
- 230000036436 anti-hiv Effects 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 7
- 108090000765 processed proteins & peptides Proteins 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- 238000004113 cell culture Methods 0.000 description 6
- 230000009089 cytolysis Effects 0.000 description 6
- 150000002500 ions Chemical class 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 230000035899 viability Effects 0.000 description 6
- 238000003149 assay kit Methods 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 5
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 5
- 210000002751 lymph Anatomy 0.000 description 5
- 230000002265 prevention Effects 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 206010018910 Haemolysis Diseases 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- 210000001772 blood platelet Anatomy 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 210000003714 granulocyte Anatomy 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000008588 hemolysis Effects 0.000 description 4
- 210000002865 immune cell Anatomy 0.000 description 4
- 230000028993 immune response Effects 0.000 description 4
- 238000001990 intravenous administration Methods 0.000 description 4
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 239000008188 pellet Substances 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- 102000004196 processed proteins & peptides Human genes 0.000 description 4
- 238000010791 quenching Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- HJHVQCXHVMGZNC-JCJNLNMISA-M sodium;(2z)-2-[(3r,4s,5s,8s,9s,10s,11r,13r,14s,16s)-16-acetyloxy-3,11-dihydroxy-4,8,10,14-tetramethyl-2,3,4,5,6,7,9,11,12,13,15,16-dodecahydro-1h-cyclopenta[a]phenanthren-17-ylidene]-6-methylhept-5-enoate Chemical compound [Na+].O[C@@H]([C@@H]12)C[C@H]3\C(=C(/CCC=C(C)C)C([O-])=O)[C@@H](OC(C)=O)C[C@]3(C)[C@@]2(C)CC[C@@H]2[C@]1(C)CC[C@@H](O)[C@H]2C HJHVQCXHVMGZNC-JCJNLNMISA-M 0.000 description 4
- 238000010561 standard procedure Methods 0.000 description 4
- 239000000758 substrate Substances 0.000 description 4
- 102000019034 Chemokines Human genes 0.000 description 3
- 108010012236 Chemokines Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000012981 Hank's balanced salt solution Substances 0.000 description 3
- 206010062016 Immunosuppression Diseases 0.000 description 3
- 102000000588 Interleukin-2 Human genes 0.000 description 3
- 108010002350 Interleukin-2 Proteins 0.000 description 3
- 208000036142 Viral infection Diseases 0.000 description 3
- 238000004820 blood count Methods 0.000 description 3
- 238000003501 co-culture Methods 0.000 description 3
- 210000003979 eosinophil Anatomy 0.000 description 3
- 229960000301 factor viii Drugs 0.000 description 3
- 238000005534 hematocrit Methods 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 230000001506 immunosuppresive effect Effects 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 210000000440 neutrophil Anatomy 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 231100000041 toxicology testing Toxicity 0.000 description 3
- 230000029812 viral genome replication Effects 0.000 description 3
- 230000009385 viral infection Effects 0.000 description 3
- AXTNFJKQZPETJA-UHFFFAOYSA-N 1-methyl ethyl 2-chloro-5-[[[(1-methylethoxy)thiooxo]methyl]amino]-benzoate Chemical compound CC(C)OC(=S)NC1=CC=C(Cl)C(C(=O)OC(C)C)=C1 AXTNFJKQZPETJA-UHFFFAOYSA-N 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 238000012286 ELISA Assay Methods 0.000 description 2
- 108010054218 Factor VIII Proteins 0.000 description 2
- 102000001690 Factor VIII Human genes 0.000 description 2
- 108010016183 Human immunodeficiency virus 1 p16 protease Proteins 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- 101100334739 Mus musculus Fgfr3 gene Proteins 0.000 description 2
- 238000011887 Necropsy Methods 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 241000700157 Rattus norvegicus Species 0.000 description 2
- 101150021948 SAM2 gene Proteins 0.000 description 2
- 101150076716 SAM3 gene Proteins 0.000 description 2
- 101150016293 SAM4 gene Proteins 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- GLWHPRRGGYLLRV-XLPZGREQSA-N [[(2s,3s,5r)-3-azido-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl] phosphono hydrogen phosphate Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](N=[N+]=[N-])C1 GLWHPRRGGYLLRV-XLPZGREQSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 238000002832 anti-viral assay Methods 0.000 description 2
- 230000005875 antibody response Effects 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 210000000702 aorta abdominal Anatomy 0.000 description 2
- 210000003651 basophil Anatomy 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000035602 clotting Effects 0.000 description 2
- 239000003636 conditioned culture medium Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000012228 culture supernatant Substances 0.000 description 2
- RMRCNWBMXRMIRW-BYFNXCQMSA-M cyanocobalamin Chemical compound N#C[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O RMRCNWBMXRMIRW-BYFNXCQMSA-M 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 235000021186 dishes Nutrition 0.000 description 2
- 238000000132 electrospray ionisation Methods 0.000 description 2
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 2
- 238000000119 electrospray ionisation mass spectrum Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000002489 hematologic effect Effects 0.000 description 2
- 230000000423 heterosexual effect Effects 0.000 description 2
- 210000005260 human cell Anatomy 0.000 description 2
- 238000010348 incorporation Methods 0.000 description 2
- 239000012678 infectious agent Substances 0.000 description 2
- 230000001524 infective effect Effects 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000004949 mass spectrometry Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000000813 peptide hormone Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000002976 reverse transcriptase assay Methods 0.000 description 2
- 238000004007 reversed phase HPLC Methods 0.000 description 2
- 238000013207 serial dilution Methods 0.000 description 2
- 238000012453 sprague-dawley rat model Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 238000003211 trypan blue cell staining Methods 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- RPZOFMHRRHHDPZ-UHFFFAOYSA-N 1-[2-(2-cyanoaziridin-1-yl)propan-2-yl]aziridine-2-carboxamide Chemical compound C1C(C(N)=O)N1C(C)(C)N1CC1C#N RPZOFMHRRHHDPZ-UHFFFAOYSA-N 0.000 description 1
- GHCZTIFQWKKGSB-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;phosphoric acid Chemical compound OP(O)(O)=O.OC(=O)CC(O)(C(O)=O)CC(O)=O GHCZTIFQWKKGSB-UHFFFAOYSA-N 0.000 description 1
- OZUBORKYZRYLSQ-UHFFFAOYSA-N 3-nitrosobenzamide Chemical compound NC(=O)C1=CC=CC(N=O)=C1 OZUBORKYZRYLSQ-UHFFFAOYSA-N 0.000 description 1
- 101710186708 Agglutinin Proteins 0.000 description 1
- 101100020619 Arabidopsis thaliana LATE gene Proteins 0.000 description 1
- 101100134902 Arabidopsis thaliana OFUT23 gene Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241001473877 Biserrula isolate Species 0.000 description 1
- 102000001902 CC Chemokines Human genes 0.000 description 1
- 108010040471 CC Chemokines Proteins 0.000 description 1
- 108010041397 CD4 Antigens Proteins 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010015719 Exsanguination Diseases 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010078851 HIV Reverse Transcriptase Proteins 0.000 description 1
- 229940033330 HIV vaccine Drugs 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 101710146024 Horcolin Proteins 0.000 description 1
- 101900297506 Human immunodeficiency virus type 1 group M subtype B Reverse transcriptase/ribonuclease H Proteins 0.000 description 1
- 229920001612 Hydroxyethyl starch Polymers 0.000 description 1
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 101710189395 Lectin Proteins 0.000 description 1
- 101000577064 Lymnaea stagnalis Molluscan insulin-related peptide 1 Proteins 0.000 description 1
- 102000009571 Macrophage Inflammatory Proteins Human genes 0.000 description 1
- 108010009474 Macrophage Inflammatory Proteins Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 101710179758 Mannose-specific lectin Proteins 0.000 description 1
- 101710150763 Mannose-specific lectin 1 Proteins 0.000 description 1
- 101710150745 Mannose-specific lectin 2 Proteins 0.000 description 1
- 101710151805 Mitochondrial intermediate peptidase 1 Proteins 0.000 description 1
- 102100030856 Myoglobin Human genes 0.000 description 1
- 108010062374 Myoglobin Proteins 0.000 description 1
- 101000737895 Mytilus edulis Contraction-inhibiting peptide 1 Proteins 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 241000282577 Pan troglodytes Species 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 108010038988 Peptide Hormones Proteins 0.000 description 1
- 102000015731 Peptide Hormones Human genes 0.000 description 1
- 241000702619 Porcine parvovirus Species 0.000 description 1
- 239000012979 RPMI medium Substances 0.000 description 1
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 1
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 1
- 206010054094 Tumour necrosis Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000000910 agglutinin Substances 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 229940121357 antivirals Drugs 0.000 description 1
- 210000000709 aorta Anatomy 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 229950009664 azimexon Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229910002056 binary alloy Inorganic materials 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 239000003139 biocide Substances 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 239000010836 blood and blood product Substances 0.000 description 1
- 229940125691 blood product Drugs 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000000701 coagulant Substances 0.000 description 1
- 238000002648 combination therapy Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000009223 counseling Methods 0.000 description 1
- 235000000639 cyanocobalamin Nutrition 0.000 description 1
- 239000011666 cyanocobalamin Substances 0.000 description 1
- 229960002104 cyanocobalamin Drugs 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 229940042399 direct acting antivirals protease inhibitors Drugs 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000003777 experimental drug Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012997 ficoll-paque Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 229940044627 gamma-interferon Drugs 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000002607 hemopoietic effect Effects 0.000 description 1
- 208000002672 hepatitis B Diseases 0.000 description 1
- 238000006897 homolysis reaction Methods 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 230000000652 homosexual effect Effects 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 229940050526 hydroxyethylstarch Drugs 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000008102 immune modulation Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 239000002955 immunomodulating agent Substances 0.000 description 1
- 229940121354 immunomodulator Drugs 0.000 description 1
- 230000002434 immunopotentiative effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229940029329 intrinsic factor Drugs 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000002433 mononuclear leukocyte Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000010807 negative regulation of binding Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 229940042402 non-nucleoside reverse transcriptase inhibitor Drugs 0.000 description 1
- 239000002726 nonnucleoside reverse transcriptase inhibitor Substances 0.000 description 1
- 239000006916 nutrient agar Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000003170 nutritional factors Nutrition 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 108010062490 p27 antigen Proteins 0.000 description 1
- 101800000007 p51 RT Proteins 0.000 description 1
- 230000000590 parasiticidal effect Effects 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 210000001539 phagocyte Anatomy 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 230000003334 potential effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000135 prohibitive effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010079 rubber tapping Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 235000010378 sodium ascorbate Nutrition 0.000 description 1
- PPASLZSBLFJQEF-RKJRWTFHSA-M sodium ascorbate Substances [Na+].OC[C@@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RKJRWTFHSA-M 0.000 description 1
- 229960005055 sodium ascorbate Drugs 0.000 description 1
- PPASLZSBLFJQEF-RXSVEWSESA-M sodium-L-ascorbate Chemical compound [Na+].OC[C@H](O)[C@H]1OC(=O)C(O)=C1[O-] PPASLZSBLFJQEF-RXSVEWSESA-M 0.000 description 1
- 239000008279 sol Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000011146 sterile filtration Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 208000010648 susceptibility to HIV infection Diseases 0.000 description 1
- 238000011285 therapeutic regimen Methods 0.000 description 1
- 239000000724 thymus hormone Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000048 toxicity data Toxicity 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 230000003253 viricidal effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2/00—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor
- A61L2/0005—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts
- A61L2/0082—Methods or apparatus for disinfecting or sterilising materials or objects other than foodstuffs or contact lenses; Accessories therefor for pharmaceuticals, biologicals or living parts using chemical substances
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/15—Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/0005—Vertebrate antigens
Definitions
- This invention relates to soluble factors having anti-HIV activity which are derived from leukocytes and immune cells of animals and humans, their method of extraction and preparation and pharmaceutical preparations comprising the anti-HIV factors.
- HIV human immunodeficiency virus infection
- AIDS Acquired Immunodeficiency Syndrome
- HIV-1 a retrovirus
- HIV-1 a retrovirus
- the virus interacts with the human immune system causing serious and oftentimes fatal immunosuppressions in infected patients.
- HIV infection is widespread throughout out society.
- the groups which run the highest risk of HIV- 1 infection include homosexual and bisexual males and intravenous drug users.
- Other high-risk groups include recipients of blood transfusions and sexual partners in the HrV-infection risk groups.
- the HIV-infection may also be transmitted merely be heterosexual intercourse. Several cases of HIV infection are known through heterosexual intercourse even where a condom was used.
- a cure for HIV-1 infection has not yet been found.
- Recent advances in HIV/AIDS research have provided valuable information which has led to the discovery of new accurate HIV diagnostic systems which detect HTV RNA measurement or "viral load", new pharmaceuticals such as protease inhibitors, and combination therapies, in addition to education and counseling. All of these efforts have contributed to better care of HTV-infected patients and disease management.
- One drug which has been approved by the Food and Drug Administration (FDA) for treatment of HTV infection is AZT (3-azido-2',3'-dideoxy-thymidine) which is believed to function by slowing or inhibiting viral replication.
- AZT does not cure the disease nor does it prevent infection.
- AZT is known to be responsible for serious side effects, such as bone ma ⁇ ow suppression, and it is poorly tolerated in a high proportion of patients. Its beneficial effects have also been reported to diminish in 12-18 months.
- anti-viral pharmaceuticals include alpha interferon, gamma interferon, ampliquen, azimexon and isopinosine.
- AZT alpha interferon, gamma interferon, ampliquen, azimexon and isopinosine.
- drugs and treatments may add a few months to a few years to the lives of HlV-infected patients who have developed AIDS, but do not cure patients of the disease. These drugs are also highly toxic and may have severe side effects.
- the HIV virus becomes resistant to these drugs and a strict dosage regimen must be maintained, with a patient taking 20 or more pills a day being a common occu ⁇ ence.
- the present invention now provides a soluble composition produced from lysate of leukocytes and immune cells from whole blood, bone ma ⁇ ow and blood and fetal cells of primates and human beings which is therapeutically effective in treating HIV-1 infection, and is also effective as a vaccine in the prevention of HIV-1 infection, as well as in the prevention of infection from several other viruses.
- the inventive composition is effective in the therapeutic reduction of viral load in infected patients, and is also effective in the treatment and prevention of other known human viruses including hepatitis virus, herpes virus, protein enveloped virus, non-enveloped virus, RNA viruses and DNA viruses; the inventive compositions are safe, non-toxic and display no adverse side reactions.
- anti-viral composition of this invention has properties of inhibition of protease activity, inhibition of reverse transcriptase activity and blockage of gpl20. This invention is more fully described by the following, detailed discussion with prefe ⁇ ed embodiments and embodiments.
- FIGs. and 1 and 2 are HPLC spectra of an active factor of the inventive composition.
- FIG. 3 is a positive ion electrospray-MS spectrum of an active factor of the inventive composition.
- FIG. 4 is an EI-MS spectrum of an active factor of the present composition.
- FIG. 5 is a "best-fit" library matter for the spectrum shown in FIG. 4.
- FIG. 6 is a UV chromatogram of an active factor of the inventive composition.
- FIGS. 7-12 are ESI-MS spectra of several major components detected within the chromatographic profile.
- FIG. 13 is a graph summarizing hematologic evaluation of whole blood after one hour contract with the inventive composition.
- FIG. 14 is a graph illustrating inhibition of HIY-l protease by composition.
- FIG. 15 is a graph illustrating inhibition of HIV-1 reverse transcriptase activity by composition.
- FIG 16 is a graph illustrating inhibition of tumor necrosis.
- Factor Alpha UI is a cell line used for analysis.
- FIG. 17 is an electrophoresis gel study of the composition of showing the difference in electropheresis pattern between human and baboon cell preparation of the composition.
- FIG. 18 is a graph showing effect of baboon PBMC extract composition on different strains of HIV- 1, chronically infected H9/IIIB cells and SIV (Simian Immune Deficiency Virus).
- FIG. 19 is a graph showing effect of baboon PBMC extract composition on pre- and post-treated human cells and inhibition of viruses with the composition, which indicates the inventive composition's efficacy as an HIV vaccine.
- FIG. 20 is a graph showing effect of baboon PBMC extract on % inhibition of HIV-1 up to 12 days of anti- viral activity at 37 °C indicating high degree of stability of the composition at elevated temperatures.
- FIG. 21 is a graph illustration results of baboon tissue culture grown cell extract composition of the present invention on reverse transcriptase activity using cell line CEM-TART cells.
- FIG. 22 is a graph illustrating results of baboon PBMC extract composition of the present invention as reverse transcriptase activity.
- CD-4 is the cell protein to which the HTV-1 virus binds.
- the protein gp 120 is the virus protein which is responsible for the binding. The virus interacts with the human immune system, with resulting immunosuppression from depletion and functional abnormalities of CD4+T cells.
- an effective method is to reduce the viral burden in the circulating blood, thereby boosting the immune system and providing essential nutritional factors to stimulate production of non- infected healthy blood cells, particularly CD4+T lymphocytes.
- composition comprising a soluble lysate product from leukocytes and immune cells of primates, such as baboons, and humans who are immune to or otherwise resist infections by viruses, microorganisms or other disease-causing agents, which is therapeutically effective in treating HIV infections and other viral infections as well as acting as a vaccine in the prevention of HIV infection and other viral infections.
- Blood, bone ma ⁇ ow, cord blood, stem cells and progenitor cells from primates, mammalians and human species may be different from other primates, mammalians and human species in genetic, enzymatic make-up, or may contain certain peculiar natural components that bestow resistance and immunity to certain viral or bacterial infections. These factors may be present in the immune cells or other blood cells or tissue that produce blood cells, and may be in the form of protein, peptides, amino acids, carbohydrates, lipids or other biochemical nature that are present naturally in the living body of these resistant-to-infections species. Blood is composed of red blood cells, platelets, stem cells, macrophages, leukocytes as cellular components.
- WBC white blood cells
- Lymphocytes in particular T cells, participate in regulating the immune system of the body, and are themselves regulated by certain peptide hormones. T cells, which control and regulate the immune system are classified as CD4+T lymphocytes and CD8+T lymphocytes. The role of other lymphocytes is not well understood, though they are known to act as phagocytes and ingest foreign particles.
- Eosinophils are known to exert potent parasiticidal effect through their cationic proteins and enzymes. Primate stem cells and granulocytes, bone ma ⁇ ow, and cord blood all secrete Viral Suppressive Factors and include Regulated Upon Activation Normal T Cells Expressed and Secreted (RANTES) Chemokines, Macrophage Inflammatory Proteins (MIP-1 Alpha and MIP-I Beta), and Beta chemokine microphage derived chemokine (MDC).
- Immunomodulators include polypeptide thymic hormone Thymosine Alpha- 1, subsets of CD4+T cells thl responsible for cellular immunity and th2 responsible for humoral immunity.
- the present inventive soluble lysate product may include by way of example, but without limitation, immunopotentiating polypeptide hormone, extracts of CD8+T lymphocytes containing viral suppressive factors, CD4+T lymphocytes containing factors that regulate cellular hormonal and immunity, hemopoietic factors, and other cell nutrient species such as calcium and ascorbic acid.
- composition of the present invention provides an active, stable and safe solution that can be sterilized and used at a physiologically active pH suitable for injection or transformation into the human body for treatment and prevention of immunosuppression diseases by immunoresponsive therapeutically active substances such as described above, and a variety of other diseases such as hepatitis B, hepatitis C, cytomegalic virus (CMV), herpes virus and other viral diseases.
- immunoresponsive therapeutically active substances such as described above, and a variety of other diseases such as hepatitis B, hepatitis C, cytomegalic virus (CMV), herpes virus and other viral diseases.
- CMV cytomegalic virus
- the inventive therapeutic composition can be prepared by first providing a leukocyte fraction of whole or cord blood derived from a primate such as a baboon, chimpanzee, monkey or gabon, which species is immune to infection from HIV and other cu ⁇ ently known human-to-human transmissible viruses.
- This leukocyte fraction may comprise one or more of stem cell, progenitor cell, lymphocytes, monocytes, and granulocytes, which contain suppressive factors and immune modulating factors.
- Peripheral blood mononuclear cells and buffy coat cells can then be separated from the leukocyte fraction using any conventional method such as the Ficol-Hypaque technique, centrifugal separation or any other method suitable for the separation of leukocytes.
- the cells are then lysed, stabilized, the pH adjusted accordingly as desired and the solution is then sterilized for therapeutic use in humans.
- the following example is illustrative of this broadly described prefe ⁇ ed embodiment.
- Nucleated cells are selectively concentrated by using procedures such as density gradient separation with hydroxyethyl starch, Ficol-Hypaque, leukocyte removal filtration, direct lysis or any other method including the use of antibody-coated bead non-erythrocyte cells from blood.
- Baboon blood is processed for obtaining buffy coat using standard protocol for isolation of PBMCs from buffy coat by Ficol-Hypaque (Histopaque) as follows:
- the purified nucleated cells are then washed 3 times with HBSS and stabilizer solution containing excess calcium ions.
- the washing is performed in a refrigerated centrifuge at 4500x g for 10 minutes at 18-20°C.
- the deposit is resuspended in HBSS without Pana Sera Plus to contain 50-500 million cells/ml.
- the cells are then ruptured by sonification under ice.
- the sonicate is repeatedly frozen and thawed by alternate immersion in liquid nitrogen and water at 37 °C in five rapid and successive cycles. After the last thaw cycle, the slurry is centrifuged in a refrigerated centrifuge at 4500x g for 10 minutes to obtain a clear supernatant.
- the supernatant is sterilized by passing through 0.2 micron membrane filter and mixed 1 : 1 by volume with Sterile Secondary Ingredients (SSI) under aseptic conditions.
- SSI comprises sterile intrinsic factor, cyanocobalamin, folic acid, sodium ascorbate, calcium chloride, potassium chloride, sodium phosphate, potassium phosphate, sodium chloride and sucrose.
- the final composition containing cell lysate and SSI has a pH of 6.8-7.4.
- the composition containing the fortified lysate of stem cells, and granulocytes is then aseptically dispensed in 1-10 ml aliquots in sterile ampules for storage in liquid nitrogen until used. Each 1 ml contains the lysate obtained from 10-50 million cells.
- THAW AND ADD STABILIZING SOLUTION (Use 10% v/v, contains Ca, and KC1 in PBS at pH 7.4)
- the inventive composition exerts a direct and indirect action on viral particles to adversely affect the replication of the infective agents.
- the composition thus lowers the rate of virus replication and virus load in the host body.
- Direct lysis of the cell-free infectious agents in the host blood is thought to be attributable to any of several viral and enzyme inhibitory factors present in the inventive lysate composition.
- transfusions with homologous blood may be necessary to restore the innate leukocyte population to its normal level in a subject host.
- the present invention also circumvents a failure of mass immunization.
- normal human BMCs bone ma ⁇ ow cells
- routine vaccines are prepared from the antigenic fragments of infectious agents. Injections of these vaccines stimulate the host to produce specific antibodies that bind to the antigen sites of the infecting organ.
- the infective organism often mutates to alter its antigenic sites so that these antibodies have no specific binding sites on the antigen molecule; under these circumstances, the humoral antibody that protected the host earlier against the parent strain of the agent becomes ineffective.
- Administration of a non-specific composition such as the one described herein, is therefore a more powerful therapeutic tool.
- inventive composition causes a reduction in viral load which is thought to potentially lead to a complete or substantially complete elimination of HIV infection in a patient.
- inventive composition also boosts the immune system by increasing the CD4 T lymphocyte count, improves conditions for new virus- free white blood cells and is non-toxic, side-effect free and completely compatible with human red blood cells.
- composition (A) described in this invention consists of:
- Composition (B) described in this invention consists of:
- composition (A) (10 ml whole blood) lysed in D. water or hypotonic solution (1.0 ml) 2.
- At least one active fraction of the present inventive composition is thought to be a relatively small molecule of molecular weight 200-500, which is refe ⁇ ed to herein Natural Killing Agent ("ANKA").
- ANKA Natural Killing Agent
- Amino acid analysis has shown that ANKA is not a protein or polypeptide type of cytokine, but is a carbohydrate. Further, Because of its small size, ANKA may be given orally, it may effectively distribute beyond the intravascular compartment and would not evoke an antibody response in humans.
- baboon whole blood in CPD as coagulant was obtained from Naval Blood Research Lab (NBRL) under sterile conditions.
- NBRL Newcastle Blood Research Lab
- blood was divided into two equal parts of 30 ml in sterile centrifuge tubes. 10 ml of histopaque was added at the bottom of the tubes.
- PBMC primate bone ma ⁇ ow cell
- the remaining PBMC fraction in the other tube was used to prepare lysate by adding 5 ml sterile water, mixing well by vortex and freezing at -20 °C for 4 days.
- Human PBMCs were provided by ABS and were used to prepare lysate simultaneously as control following same procedure as for baboon cell and lysed in 5 ml of sterile distilled water and frozen for 4 days.
- Frozen lysates were thawed and examined under microscope for cell debris, DNA and other substances. Both human and baboon cells were completely lysed leaving no intact cells in the solution, cell debris and DNA strands were seen in both preparations.
- Both tubes were centrifuged at 1500 G for 10 minutes, and supernatant was filtered using 0.22 micron filter. Both tubes gave clear supernatant and were frozen at -20 °C.
- both tubes were thawed, examined for turbidity, centrifuged at 1500G for 15 minutes; no precipitate was seen and the solutions were aliquoted in 100 ul fractions and frozen.
- 0.3 ml baboon preparation was diluted in 30 ml sterile saline and saved.
- Tissue culture was terminated with cell count over 200 million, the cells washed twice with PBS and lysed with 14 ml distilled water, complete lysis was observed after few minutes. The lysate was stored at -18°C.
- Baboon cell preparation contained 1.4 gm total protein per ml.
- FIGs 1 and 2 are HPLC spectra of an apparent active factor of the inventive composition.
- the compound is thought to be of a relatively small molecular weight containing a benzene ring, possibly with an alkyl substituent.
- the spectra is characteristic of phenylalanine, with the compound possibly being a metabolite or antibiotic that is in cells.
- Electrospray ionization spectrometry was performed on the isolated substance having a peak at 215 nm in HPLC analysis using a VG Biotech BIO-Q instrument with a quadruple analyzer. Myoglobin was used to calibrate the instrument. Sample aliquots of 50 ul were injected into the instrument source. Elution was carried out using a mixture of 50% acetonitrile containing 0.1% TFA, at a flow rate of lOul/min.
- this sample gave a positive ion electrospray-MS spectrum showing notable possible sample-related signals at m/z 86, 194 and 288.
- the appearance of this spectrum with the absence of no readily assignable pseudomolecular ions(s) may suggest this sample to not be peptide/protein in nature, or a non-nitrogen containing analyte with a lipid-like structure.
- Electron Impact Mass Spectrometry was carried out on M-Scans VG analytical ZAG 2-SE high field mass spectrometer with reference to SOP# MS-003, MS-002 and MS-007.
- the source temperature was maintained at 200 °C and spectra were acquired with an electron voltage of 70eV.
- the sample was inserted into the instrument via the solids probe and heated until an appropriate spectrum obtained. Calibration was performed using PFK.
- UV detection was achieved using an HP variable wavelength detector operating at 215 nm.
- MS detection was achieved using a VG BioQ triple quadruple mass spectrometer operating in the positive ion electrospray ionization mode, with the following parameters: 1. Scan Range : m/z 35-700 2. Cone Voltage : 57V
- FIG. 4 illustrates a typical EI-MS spectrum obtained during analysis of the sample. This shows a number of major possible sample-related signals including those at m z 69, 87, 114, and 129.
- FIG. 5 illustrates a "best-fit" library match for this spectrum and although not identical, is similar to that of dimethyl decane.
- FIG. 6 illustrates the UV chromatogram obtained following sample injection. In contrast to previous analyses, this shows a major component eluting at 26.74 min. together with numerous additional components eluting throughout the gradient.
- FIGS. 7-12 illustrate the ESI-MS spectra obtained for a number of the major components detected within the chromatographic profile. Most of these appear to show distinct, strong possible (M+H) + at pseudomolecular ions which are summarized in Table 3 below. TABLE 3: Retention time and possible m/z value for the pseudomolecular ions detected during LC-MS
- the major peak (26.74 min.) shows a possible (M+H) + at mz/ 383.6 which has not been detected in previous analyses.
- Peaks at 32.42 and 36.93 min. show several major signals with a 44amu differences. This is indicative of the present of an ethoxylate polymer.
- 1/10 dilution has 210 ug 1/100 dilution has 21 ug 1/500 dilution has 4.25 ug 1/1000 dilution has 2.1 ug 20 1/2000 dilution has 1.0 ug
- BMCs from 10.0 ml freshly drawn heparinized human whole blood were isolated using the standard BBI protocol. BMCs were diluted in 10 ml of RPMI- 1640 tissue culture media (see SIGMA catalog, pp. 1473-94), mixed well by gentle inversion and divided into two equal parts in 15 ml tubes. 0.1 ml of 1/10 composition was added to one of the tubes and labelled as test (treated), and the other untreated tube was used as control.
- Composition was first added to plasma at 1:10 dilution, and samples were incubated for 1, 2, 5, and 23 hours at 37°C, after which they were frozen and assayed for Factor VIII and Factor IX clotting activity by one stage APTT method. 0.15M sodium chloride was used as control. Samples were also incubated at room temperature and at 4°C.
- Protocol for preparation of infected cells in HIV-1 inhibition test can be summarized below as follows. Normal PBMCs (from Red Cross) were infected with 6-log HIV-1 MN strain
- PMBCs 2x10 -5 were incubated with HIV-1 MN at 35 °C for 3 hours in a 15ml tube.
- the excess free virus was removed by washing the cells with lx PBS 2 times.
- the infected PBMCs were cultured in RPMI + 10% bovine calf serum + PS at 37°C and 5% CO 2 .
- the blood was divided in four aliquots 12 ml each.
- PBMCs were isolated using BBI protocol for Ficol-Hypaque isolation of PBMCs (standard procedure).
- each tube was added to one infected set. Then they were transfe ⁇ ed to 3 rows of a 24 well plate.
- PBMCs undiluted 1:10 1 :100 1 :1000 1 :10000 1 :1000000 uninfected - - - - and - - - - treated - - - - -
- the plate was incubated for 7 days at 37 °C with 596 CO 2 .
- Fresh blood was added (196) to 10 ml PBS and treated with 100 ul of baboon lysate at RT. No hemo lysis was observed after 2 weeks.
- Dav 4 Use 100 ul of stabilizing solution to add to 1 ml PBMCs lysate.
- PBMCs Normal blood was drawn and PBMCs were isolated using standard protocol. The PBMCs were counted and cultured in RPMI. Dav 5
- Dupont P-24 Kit was used to determine the P-24 output of all samples, except for the negative control where only 2 tubes were used.
- HIV-1 MN HIV reagent program CAD # 137
- RPMI media Difco
- HIV-1 MN HIV-1 MN (AIDS reagent program CAT # 137)
- PBMCs isolated as per BBI standard protocol and incubated with HIV-1 MN at 37 C for 6 hours in 15mL tube. The excess (free viruses) were removed by washing the cells PBS one time.
- Test for Virucidal Activity, Cell Viability Toxicity to Red Cells, Etc. as follows: a. Raw 10 ml fresh human blood in heparinized syringe and transfer to 50 ml centrifuge tube. b. Isolate PBMC as above in step 3 and use these human PBMC as indicator cells. c. Set up 4 wells as controls, add 100 ul indicator cells and 6 log HIV virus in tissue culture media. d. Set up remaining wells for testing the drug sensitivity to viruses, add 100 ul indicator cells and 6 log HIV virus in tissue culture media. e. Incubate 37°C for 7 days. Check syncytia formation after 24 hours, 3 days, 7 days. f.
- Tissue culture grown cells are equally potent as viral inactive agent as fresh baboon cells.
- PBMC fraction from both baboons as well as human PBMC fractions were added to the culture media for growing PBMC fractions in tissue culture and cells were examined under microscope on daily basis for 7 days.
- PBMC lysates were made by adding cells to 1.0 ml distilled water and storing the cells at 4°C overnight. 100 ul stabilizing solution was added to all lysates and lysates were tested for virus inactivation as above.
- Each lysate was tested at dilutions 1/10, 1/50, 1/100, 1/200, 1/400, and 1/800. 750ul of each dilution of the lysate was added to the test wells containing HIV (6 log) in 50 ul media, human PBMC in 200 ul culture media.
- Control wells contained no drug. 750 ul culture media was added to make the volume.
- HIV-1 MN HIV-1 MN (AIDS reagent program CAT #137
- Lysate 1 (# 266 cells) (infected PBMCs) (composition treated)
- Lysate 3 (# 266 & 267 tissue culture grown cells pooled)
- Lysate 4 (# 267 tissue culture grown cells)
- Lysate 5 (Lysate from baboon # 1-3/29/96 tested for stability and reproducibility)
- Freshly obtained human and baboon blood (15 ml each) were diluted with equal volume of Hanks balanced salt solution (final 30-ml) and carefully laid over 10 ml of Ficoll- Paque (Sigma). Samples were centrifuged at 550 x g at 20°C for 40 min. Top layers were carefully discarded and middle layers containing PBMC were collected. The cells were washed twice with PBS (10 ml each) followed by centrifugation at 400 x g at 20 and 10°C in the first and second wash respectively. Cells were then re-suspended in 10 ml RPMI- 1640 (serum-free) for cell count and viability (3 x 10 7 PBMC/10 ml).
- HIV reverse transcriptase assay was performed in duplicate by incorporation of 32P-dttp into oligo-A template primed with oligo-dt. In this experiment CEM-TART cells were treated overnight with composition. Followinged by 6 days infection with HIV-1 after-rev clone. Reverse transcriptase activity was measured after detergent treatment of the cell supernatants according to the established methods. RESULTS
- composition at a concentration of 1 :500 consistently resulted in 60-70% inhibition of HIV growth as measured by reverse transcriptase assay. See FIG. 21. Inhibitory activity of composition at a higher than 1 :500 dilution was not significant. 3H-thymidine incorporate assay (indicative of cell growth) was also inhibited up to 30% by 1 :500 diluted composition. Results are summarized below in Tables 6 and 7.
- R 2 0 88 0 01 0 01 043 020
- EXAMPLE 10 L97-85 BABOON CELL EXTRACT ANTI-HIV ACTIVITY
- PBMC peripheral blood mononuclear cells
- the stabilized lysate was diluted to 50 ml in saline, and aliquot in 1ml vials for storage and future use.
- Cell extract was diluted used with equal volume of HIV-1 IIIB dilution to infect human PBMC overnight. Cells were then washed and placed in culture in media containing indicated dilution of the composition. 1/2 media changes with media containing the indicated dilutions were made at 3 day intervals. Conditioned media was tested by HIV-l p 27 antigen capture. The results are summarized below.
- Target cells were activated human PBMCs and were incubated with the indicated virus at 1 : 10 dil. and dilution of extract overnight. Cells were then washed and plated in the indicated dilution of extract with 1/2 fluid changes at 3-4 day intervals. Antigen capture was with HIV p25 OTC kit. This assay is summarized below in Table 11.
- the object of this assay is to determine whether a drug will inhibit HIV-1 protease activity. This assay is summarized below as follows: Materials: l. ENZYME
- the HTV-1 Protease is received from Bachem Bioscience, Inc. (H-1256) at a concentration of 0.116 mg/ml. Thaw the stock, aliquot into 15 ul samples and refreeze at -70 °C. Dilute the stock to a working concentration 1.25 ug/ml in IX buffer.
- Armide is prepared by multiple Peptide Systems, as a dry powder. Dilute the 10 mM stock in ddH 2 0 (MW of peptide is 1044 and the stock is 10.44 mg/ml). Dilute the stock to a lmM working solution in IX buffer.
- QUENCH SOLUTION Prepare a 9:1 mix of 8 M Quanidine-HCL to 10:TGA. Use 40 ul of this mix to stop the assay.
- PLATE ANALYS IS REPORT .P ACE : 1 _:
- the object of this study was to investigate the potential effects of composition during intravenous injections (48) hours apart) followed by a recovery period for a total of 28 days.
- the dose level was selected by the Sponsor according to the potential human exposure, existing toxicity data and any limitations imposed by the test article.
- test article was used as supplied by the Sponsor and administered by intravenous injection, via the tail vein on 5 occasions (48 hours apart on Days 1, 3, 5, 7 and 9) and the final dose was administered on the day of necropsy (Day 28).
- the dose volume administered to each animal was 1 mL/occasion.
- BLOOD SOURCE aorta, orbit, tail vein
- Eosinophils (Eosin) % 123 1 1 0 - 3
- Basophils (Baso) % 123 0 0 0 - 0
- Step 1 Add composition to
- Step 2 Mix, then incubate at room temperature for 1-6 hours.
- Step 3 Centrifuge and wash three times with equal volume of phosphate buffer saline.
- Step 4 Storage life:
- HIV-l from infected SUPT-1 T cells was co-cultured to uninfected SUPT-1 cells 48 hours prior to composition treatment. After 1 hour of treatment with 10% or 20% composition at 37°C, composition was removed by washing. Virus replication was measured by determination of HIV-l P-24 in the supernatants after 5 days in co-cultures. Cell viability was determined by Trypan Blue Dye Exclusion of pooled cells at culture termination.
- Tables 21 through 23 below show the effects of the composition on cell viability and human red blood cell parameters.
- Table 21 shows cell viability
- Table 22 shows hematological parameters
- Table 23 shows effect on red cell fragility.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Hematology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Developmental Biology & Embryology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
Claims
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BR9907991-7A BR9907991A (en) | 1998-01-30 | 1999-01-29 | Sweetening agent |
AU24820/99A AU2482099A (en) | 1998-01-29 | 1999-01-29 | Blood cell composition as a vaccine for hiv and other pathogenic organisms |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US1583098A | 1998-01-29 | 1998-01-29 | |
US09/015,830 | 1998-01-29 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO1999038520A1 WO1999038520A1 (en) | 1999-08-05 |
WO1999038520A9 true WO1999038520A9 (en) | 1999-10-21 |
Family
ID=21773878
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US1999/001922 WO1999038520A1 (en) | 1998-01-29 | 1999-01-29 | Blood cell composition as a vaccine for hiv and other pathogenic organisms |
Country Status (2)
Country | Link |
---|---|
AU (1) | AU2482099A (en) |
WO (1) | WO1999038520A1 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NO316878B1 (en) * | 2001-09-07 | 2004-06-07 | Næss Andresen Carl Fredrik | Isolated and substantially purified compound, method, pharmaceutical composition and use |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5610035A (en) * | 1983-12-05 | 1997-03-11 | Institut Pasteur Centre National De La Recherche Scientific | Methods for the preparation of hybridomas producing lymphadenopathy-associated virus (LAV) GP110-specific monoclonal antibodies and methods for the purification of GP110 employing said monoclonal antibodies |
-
1999
- 1999-01-29 WO PCT/US1999/001922 patent/WO1999038520A1/en active Application Filing
- 1999-01-29 AU AU24820/99A patent/AU2482099A/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
AU2482099A (en) | 1999-08-16 |
WO1999038520A1 (en) | 1999-08-05 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Söderhäll et al. | The cytotoxic reaction of hemocytes from the freshwater crayfish, Astacus astacus | |
KR0162094B1 (en) | Platelet membrane microparticles | |
JPH03503287A (en) | Virus-inactivated albumin suitable for living organisms | |
EP0318562A1 (en) | Method of selectively inhibiting hiv. | |
CA2165065A1 (en) | System for viral inactivation of blood | |
US5326788A (en) | Biological fluid purification system | |
US5565200A (en) | Pharmaceutical preparations derived from korean mistletoe | |
CN1683522A (en) | Whole blood quality control substance as cell bio-activity protector and its preparing method | |
CA1063019A (en) | Extracts of the haemopoietic system | |
JPH03504815A (en) | blood purification system | |
US5547674A (en) | Pharmaceutical preparations derived from European mistletoe | |
US6235239B1 (en) | Virucidal and bactericidal agent for use in the disinfection of biological fluids | |
WO1999038520A9 (en) | Blood cell composition as a vaccine for hiv and other pathogenic organisms | |
Courreges et al. | In vitro antiphagocytic effect of Melia azedarach leaf extracts on mouse peritoneal exudate cells | |
Miller et al. | Thymic dysplasia (“Swiss agammaglobulinemia”): II. Morphologic and functional observations | |
Ayoade et al. | Effects of aqueous Colocasia esculenta extracts on selected biochemical parameters in phenyl hydrazine induced male anemic albino rats | |
Karrow et al. | Thalidomide stimulates splenic IgM antibody response and cytotoxic T lymphocyte activity and alters leukocyte subpopulation numbers in female B6C3F1 mice | |
Monari et al. | Monocyte dysfunction in patients with acquired immunodeficiency syndrome (AIDS) versus Cryptococcus neoformans | |
US20050159390A1 (en) | Treatment and prevention of HIV and other viral infections | |
Mansour et al. | Effect of cyclophosphamide treatment on the response of rat peripheral blood lymphocytes to phytohemagglutinin | |
US6303154B1 (en) | Chemical alteration of mammal urine and mammal blood | |
Pitts | Con A cytotoxicity: a model for the study of key signaling steps leading to lymphocyte apoptosis in AIDS? | |
JP2002533400A (en) | Treatment and prevention of HIV and other viral infections | |
CN1051451C (en) | Anti-AIDS-virus drug composition | |
EP1555026A2 (en) | Chemical modification of mammalian blood |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
AK | Designated states |
Kind code of ref document: C2 Designated state(s): AL AM AT AU AZ BA BB BG BR BY CA CH CN CU CZ DE DK EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT UA UG US UZ VN YU ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: C2 Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
COP | Corrected version of pamphlet |
Free format text: PAGES 1-71, DESCRIPTION, REPLACED BY NEW PAGES 1-71; PAGES 72-77, CLAIMS, REPLACED BY NEW PAGES 72-77; PAGES 1/22-22/22, DRAWINGS, REPLACED BY NEW PAGES 1/22-22/22; DUE TO LATE TRANSMITTAL BY THE RECEIVING OFFICE |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
122 | Ep: pct application non-entry in european phase |