CN102671193A - Herpes simplex virus II type gene recombined attenuated live vaccine and preparation method thereof - Google Patents
Herpes simplex virus II type gene recombined attenuated live vaccine and preparation method thereof Download PDFInfo
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- CN102671193A CN102671193A CN2012101593255A CN201210159325A CN102671193A CN 102671193 A CN102671193 A CN 102671193A CN 2012101593255 A CN2012101593255 A CN 2012101593255A CN 201210159325 A CN201210159325 A CN 201210159325A CN 102671193 A CN102671193 A CN 102671193A
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Abstract
The invention discloses a herpes simplex virus II type gene recombined attenuated live vaccine and a preparation method thereof. A UL43-UL47 gene in a gene group is removed. A blister solution of herpes labialis and herpes progenitalis patients with obvious blisters is used for separating and identifying wild type HSV-1 (Herpes Simplex Virus-1) and HSV-2; after the HSV-1 and the HSV-2 are subjected to enlarged cultivation, a whole virus gene group is extracted; Vero cells are commonly transfected on shuttle vectors with isogeny side wing sequences of 700-2000 bp at left and right sides of a fluorescence-containing expression gene and a simulated and removed HSV gene; one fluorescent protein gene or a plurality of types of the fluorescent proteins are used under a fluorescence microscope as a mark/marks to select a recombinant virus; and the recombinant virus is subjected to plaque purification to obtain the needed recombined attenuated live vaccine. The vaccine can be used for effectively inducing organism mucosa and body fluid to be subjected to immune response, so as to reduce the morbidity of crowds; and a cell immune response is activated so that an infected virus can be cleaned, and the recurrence and transmission of herpes are controlled.
Description
Technical field
The invention belongs to the biological medicine technology field, be specifically related to a kind of herpes simplex virus I I type of can preventing and treating and infect gene recombinaton attenuated live vaccine of associated diseases and preparation method thereof.
Background technology
Herpes simplex virus (herpes simplex virus; HSV) be a kind of virus through skin and the close contact transmission of mucosa; Comprise herpes simplex virus I-type (being called for short HSV-1) and herpes simplex virus I I type (being called for short HSV-2); Two kinds of viruses have the collaborating genes characteristic more than 83%, the dna homolog property more than 50%.Wherein the I type can be up to more than 80% at crowd's infection rate; And hide for a long time in sensory ganglion; Though most of HSV-1 the infecteds do not have manifest symptom, minority the infected can produce lifelong very high ophthalmic ophthalmia and the fatefulue herpetic pneumonia and the herpesencephalitis of face mouth herpes, blinding property of outbreak repeatedly.HSV-2 is a primary etiological agent of herpes genitalis, accounts for more than 90% of genital herpes primary infection.Mainly cause reproductive system constitutional and recurrent herpes behind the II herpes simplex infections mankind, can hide throughout one's life in neuroganglion, recurrence when immunity of organisms reduces.The anemia of pregnant woman is prone to miscarry after infecting HSV-II type virus, also can cause fetus congenital malformation or mental retardation.Neonate infects when childbirth can cause hyperpyrexia, dyspnea or central nervous system pathological change, and wherein the neonate of 60%-70% maybe be dead.In addition, there are some researches show that genital herpes can increase 2-3 doubly with the HIV infection risk, and promote the propagation of HIV.Research shows effectively prophylaxis of herpes viral infections of condom, and world wide internal genitalia herpes sickness rate is rising year by year, according to U.S.'s recent statistics data show; U.S. adult genital herpes prevalence has risen to more than 16%; Risen to more than 25% in New York, do not had herpes simplex infections at present, especially the specific treatment method of recurrent infection; Though the antiviral drugs routine administration can be alleviated some clinical symptoms; But can not thoroughly remove the virus of hiding in the body, can not reduce recurrent number, therefore press for the vaccine that exploitation can effectively prevent and treat herpes simplex infections.
Nearly recent decades, the result of study of countries in the world HSV vaccine showed that a little less than traditional inactivated vaccine and the subunit vaccine immunogenicity, only can induce HI, cellular immunization induces effect relatively poor; Though the Novel DNA vaccine can be induced body fluid and cellular immunization simultaneously, but its transfection efficiency is lower, the effective mucosal immune response ability of through mucous membrane immune induction is lower.And in the immunne response process of herpes simplex virus; Cellular immunization and mucosal immunity are being brought into play bigger effect; But above-mentioned several types of vaccines are effectively inducing cell immunity and mucosal immune response all, therefore, and up to the present; Still do not have the vaccine listing that can use clinically, HSV vaccine all is in the clinical preceding or clinical research stage basically.
The virus attenuated live vaccine is the most classical vaccine form; Such vaccine such as antismallpox vaccine, poliomyelitis vaccine etc. are resisted the mankind on the history of infectious disease and have been brought into play irreplaceable effect, for human beings'health has been made indelible contribution.Attenuated live vaccine comprises whole or most of antigens of virus basically; And have the same infection of wild-type virus and duplication characteristic; But do not have pathogenic effects, this type of vaccine can induce body to produce comprehensive immunne response effectively, for the vaccine of other kind; Immunogenicity is strong, protects effective.
Traditional attenuated live vaccine mainly obtains through continuous culture screening, but this method time and effort consuming, is difficult to obtain the herpes simplex virus strain of pathogenic reduction in this way.In recent years, researcher begins to adopt modern molecular biology technique, and the genomic specific gene of HSV is knocked out to obtain recombinant strain.The present invention utilizes modern molecular biology technique; With HSV-1 among the Genebank and HSV-2 genome reference sequences (HSV-1 accession number: NC_001806; HSV-2 accession number: NC_001798) be source, knock out a plurality of genes, filtered out HSV-1 type and the recombinant attenuated strain of HSV-2 type through uniting; Reduction has in various degree all taken place in their pathogenecity, can infect associated diseases in order to prevent and treat HSV-1 and HSV-2 effectively.
Summary of the invention
The purpose of this invention is to provide and to prevent and to treat herpes simplex virus I I and infect the recombined attenuated live vaccine of associated diseases and the method for preparing of said vaccine.
The herpes simplex virus I I type recombinant strain that the present invention relates to gets through reorganization, screening and purification by wild HSV-2.
The technical scheme that is adopted is comparatively tangible herpes labialis of water intaking bubble and genital herpes patient's a BF, and isolation identification goes out wild type HSV-1 and HSV-2; Extract viral complete genome group after the amplification culture; With contain the luciferase expression gene and draft the common transfection Vero of the shuttle vector cell of the homologous flanking sequence of the HSV gene left and right sides 700bp to 2000bp that knocks out; Under fluorescence microscope, use a kind of or unite and use multiple fluorescence protein gene to serve as a mark to pick out recombinant virus, through obtaining needed recombined attenuated live vaccine behind the plaque purification.
Can obtain a kind of herpes simplex virus I I type gene recombinaton attenuated live vaccine, characteristic is the HSV-2 recombinant strain that has knocked out UL43, UL44, UL45, UL46 and UL47 gene in its genome.
Can obtain a kind of aforesaid herpes simplex virus I I type gene recombinaton attenuated live vaccine, be to unite UL43～UL47 gene of having knocked out in its genome and duplicate with other or infect the segmental HSV-2 recombinant strain of dispensable gene: said other duplicates or infect the dispensable gene fragment is US9～US12 gene.
On the basis of above-mentioned recombinant strain, can further knock out other obtains pathogenic further attenuating like gene recombinant strain with similar method.
It is provable on one's body to act on mice, and above-mentioned recombinant strain virus pathogenecity all has weakening in various degree, can protect mice to avoid the attack of lethal dose herpes simplex virus I-type or II type; Act on Cavia porcellus and prove that on one's body recombinant strain can prevent and treat the Cavia porcellus genital herpes.Above-mentioned recombinant strain can be used as the recombinant herpes simplex virus attenuated live vaccine with relevant disease due to the control herpes simplex infections.
Available from Stragene company, sequence and physical map are seen the catalogue of the said firm in order to the pShuttle-CMV plasmid that makes up the homologous recombination shuttle plasmid in the present invention.
The used fluorescent labeling gene of the present invention is egfp expression box and red fluorescent protein expression cassette.
Beneficial effect of the present invention: compare traditional vaccine; The attenuated live vaccine of the present invention's preparation all is through the attenuated live vaccine after the genetic engineering reorganization; Being difficult for taking place virulence replys; Other biological characteristics is similar with the wild type herpes simplex virus, but only can stimulate body generation immunne response and not have pathogenic effects after getting into body, has good safety; This vaccine has kept most of antigen of herpes simplex virus I I type; Have higher immunogenicity, can simulate wild type HSV-2 well, induce body mucosa and HI effectively; Make up the first line of defence of body opposing herpes infection, reduce crowd's sickness rate; The activated cell immunne response, infective virus might be removed in the defence line, second road of structure body, the recurrence and the propagation of control herpes.
Description of drawings
The construction strategy sketch map of Fig. 1 recombinant herpes simplex virus attenuated live vaccine
Fig. 2 recombinant herpes simplex virus attenuation II type live vaccine JSH02L structural representation
Fig. 3 recombinant herpes simplex virus attenuation II type live vaccine JSH02LS structural representation
The specific embodiment
The overall construction strategy of herpes simplex virus I I type recombined attenuated live vaccine is referring to accompanying drawing 1 in the technical scheme of the present invention.Extract wild type HSV-1 and HSV-2 complete genome group; Structure contains green or red fluorescence expressing gene and drafts the homologous recombination shuttle vector of the flanking sequence of the HSV gene left and right sides 700bp to 2000bp that knocks out; The two common transfection Vero cell; Under fluorescence microscope, select the recombinant virus of performance respective color fluorescence, through obtaining to have knocked out HSV-1 or HSV-2 recombined attenuated live vaccine behind the plaque purification.Wherein:
Knocked out the HSV-2 recombinant strain of HSV-2UL43, UL44, UL45, UL46 and UL47 gene, called after JSH02L;
Knocked out the HSV-2 recombinant strain of HSV-2UL43, UL44, UL45, UL46 and UL47 and US9, US10, US11 and US12 gene, called after JSH02LS.
On the basis of above-mentioned recombinant strain, can further knock out other obtains pathogenic further attenuating like gene recombinant strain with similar method.
Embodiment 1 recombinant herpes simplex virus II type attenuated live vaccine JSH02L makes up
1. be that HSV-1 genome method for distilling extracts the HSV-2 genome among 201010253601.5 the embodiment 1 according to the former number of patent application of this case.
2. make up homologous recombination shuttle vector pShuttle-02L-GFP.
(1) going up accession number with reference to NCBI is the sequence of the genomic UL43-UL47 of HSV-2 of NC_001798, designs the amplimer of right side flanking sequence of left side flanking sequence and the UL47 gene of UL43 gene respectively.
(2) design of primers of UL43 flanking sequence
In 5 ' terminal NotI site and three protection bases of adding of UL43 left side flap sequence forward primer, 5 ' terminal PmeI, AseI site and three protection bases of adding at UL43 left side flap sequence downstream primer reclaim the purpose band after 30 circulations of pcr amplification.
(3) design of primers of UL47 flanking sequence
UL47 right side flap sequence forward primer 5 ' terminal PmeI and MluI site and three protection bases of adding, UL47 right side flap sequence downstream primer 5 ' terminal HindIII site and three protection bases of adding reclaim the purpose band after 30 circulations of pcr amplification.
(4) the GFP expression cassette on the pcr amplification GFP expression plasmid pLKO-GFP (making up) by our company oneself; Forward primer 5 ' terminal AseI site and three protection bases of adding; 5 ' terminal PmeI site and three protection bases of adding of downstream primer reclaim the purpose band after 30 circulations of pcr amplification.
(5) cut the PCR product of GFP with AseI and PmeI, reclaim subsequent use.
(6) with NotI and PmeI enzyme action UL43 flanking sequence PCR product, PmeI and HindIII enzyme action UL47 flanking sequence PCR product reclaim the enzyme action product.
(7) in two steps with step 2.5) and 2.6) in product cloning to the pShuttle-CMV carrier that reclaims promptly make up successful pShuttle-02-GFP.
3. homologous recombination is produced HSV-2 and is knocked out strain JSH02L
(1) extracts the pShuttle-02L-GFP plasmid with the little extraction reagent kit of Axygen plasmid by its description.
(2) with liposome 2000 with HSV-2 genome and pShuttle-02L-GFP plasmid co-transfection to the vero cell.
(3) under inverted fluorescence microscope with the cell sucking-off at green CPE place, be transferred in the EP pipe that contains the 1ML culture medium.
(4) with EP pipe-80 ℃ and 37 ℃ of multigelations three times.
(5) with viral dilution to suitable titre, carry out the plaque purification with the low melting-point agarose cladding process, under fluorescence microscope, choose green plaque, carry out 5 take turns the plaque purification after acquisition HSV-1 knock out strain JSH02L (Fig. 2).
(6) with using TCID behind the Vero cell proliferation
50Method is measured titre.
Embodiment 2 uses green and the red fluorescence labelling makes up HSV-2 recombined attenuated live vaccine JSH02LS
The former number of patent application of the similar this case of the construction strategy of JSH02LS and method is the construction method of 201010253601.5 previous JSH01LS, and specifically details are as follows:
1. make up US9, US10, US11 and US12 gene and unite the HSV-2 recombinant strain JSH02S that knocks out.
(1) makes up homologous recombination shuttle vector pShuttle-02-RED.
1.1) to go up accession number with reference to NCBI be the sequence of the genomic US of the HSV-2 district US9-US12 of NC_001798, designs the amplimer of right side flanking sequence of left side flanking sequence and the US12 gene of US9 gene respectively.
1.2) design of primers of US9 flanking sequence
In 5 ' terminal NotI site and three protection bases of adding of US9 left side flap sequence forward primer, 5 ' terminal PmeI, AseI site and three protection bases of adding at US9 left side flap sequence downstream primer reclaim the purpose band after 30 circulations of pcr amplification.
1.3) design of primers of US12 flanking sequence
US12 right side flap sequence forward primer 5 ' terminal PmeI and MluI site and three protection bases of adding, US12 right side flap sequence downstream primer 5 ' terminal HindIII site and three protection bases of adding reclaim the purpose band after 30 circulations of pcr amplification.
1.4) with BglII and BamHI double digestion pDsred1-c1, connect, transform, get the pDsredbb plasmid.
1.5) cut the pDsredbb plasmid with AseI and MluI, electrophoresis reclaims the 1.5kb fragment
1.6) with NotI and PmeI enzyme action US9 flanking sequence PCR product, PmeI and HindIII enzyme action US12 flanking sequence PCR product reclaim the enzyme action product.
1.7) in two steps with 1.5) and 1.6) promptly make up successful pShuttle-012-RED behind product cloning to the pShuttle-CMV carrier that reclaims in the step.
(2) homologous recombination is produced HSV-2 and is knocked out strain JSH02S
2.1) extract the pShuttle-02L-RED plasmid with the little extraction reagent kit of Axygen plasmid by its description.
2.2) with liposome 2000 with HSV-2 genome and pShuttle-02L-RED plasmid co-transfection to the vero cell.
2.3) under inverted fluorescence microscope with the cell sucking-off at red CPE place, be transferred in the EP pipe that contains the 1ML culture medium.
2.4) with EP pipe-80 ℃ of-37 ℃ of multigelations three times.
2.5) with viral dilution to suitable titre, carry out the plaque purification with the low melting-point agarose cladding process, under fluorescence microscope, choose red plaque, carry out 5 take turns the plaque purification after acquisition HSV-2 knock out strain JSH02S.
2.6) subsequent use with extracting genome behind the Vero cell amplification JSH02S.
2. make up HSV-2 recombined attenuated live vaccine JSH02LS----homologous recombination production HSV-2 and knock out strain JSH02LS
(1) extracts the pShuttle-02L-GFP plasmid with the little extraction reagent kit of Axygen plasmid by its description.
(2) with liposome 2000 with JSH02S genome and pShuttle-02L-GFP plasmid co-transfection to the vero cell.
(3) under inverted fluorescence microscope with the cell sucking-off at green CPE place, be transferred in the EP pipe that contains the 1ML culture medium.
(4) with EP pipe-80 ℃ and 37 ℃ of multigelations three times.
(5) with viral dilution to suitable titre, carry out the plaque purification with the low melting-point agarose cladding process, under fluorescence microscope, choose green plaque, carry out 5 take turns the plaque purification after acquisition HSV-2 knock out strain JSH02LS, referring to accompanying drawing 3.
Embodiment 3 recombinant herpes simplex virus II type live vaccine are to the immunoprotection experiment of mice
1. experiment material:
Wild type herpes simplex virus I I type and recombinant herpes simplex virus attenuated live vaccine are with above-mentioned embodiment.
The normal cell lysate: the Vero cell of normal cultured scrapes the back at-80 ℃ and 37 ℃ of multigelation after-filtration in the culture dish.Common grade experiment white mice: 50, female, body weight 20 native 2g; Each is only all in the different parts marked of its health; Respective markers is being converted into 1-50 totally 50 Arabic numerals, is being divided into 10/group, dividing negative group, JSH02L, JSH02LS group by randomization.
2. immunity and counteracting toxic substances scheme
Immunizing dose: 10
6The TCID50 recombinant virus; Approach: nasal cavity instils, and promptly challenge trial was carried out with wild type HSV-2 in (35 days) the 5th week, calculated mouse death rate, confirmed to protect effect.
3. operating procedure:
(1) after the mice random packet, negative control group via intranasal application instillation approach awards 50 microlitre normal cell lysates, and other group awards 50 microlitre JSH02L, JSH02LS respectively, observes the mice situation.
(2) the 5th weeks, the counteracting toxic substances experiment;
1) draws 0.1ml HSV-2 virus with the 1mL syringe and (contain 10
6IU).
2) injection HSV-2 0.1ml in the mouse peritoneal.
3) observe the mice situation
4, result: the 5th day begins to occur death condition behind the negative control group mice counteracting toxic substances, and the 13rd back is all dead, all survivals of other group.Other group and negative group significant difference, vaccine can be resisted the lethal dose virus attack.
Embodiment 4 recombinant herpes simplex virus II type live vaccine are to the immunoprotection experiment of Cavia porcellus genital herpes
1, experiment material: wild type herpes simplex virus I I type: with measuring titre behind the Vero cell amplification HSV-2, titre is about 1 * 10
7IU/ml; Recombinant herpes simplex virus attenuated live vaccine JSH02L, JSH02LS: be about 1 * 10 with behind the Vero cell amplification titre all being adjusted
7IU/ml;
The common grade experimental guinea pig: 50, female, body weight 275 native 5g; Each is only all in the different parts marked of its health; Respective markers is being converted into 1-50 totally 50 Arabic numerals, is being divided into 10/group, be divided into matched group, JSH02L, JSH02LS group by randomization.
2, immunity and counteracting toxic substances scheme
Immunizing dose: 10
5TCID
50Live recombined vaccines; Approach: nasal cavity instils, and implements initial immunity in 0 week, and 2 weeks (14 days) are carried out booster immunization once later, and after two weeks, promptly challenge trial was carried out with wild type HSV-2 in (35 days) the 5th week, calculated Cavia porcellus pudendum disease scores accumulated, confirmed to protect effect.
3, operating procedure:
In (1) the 0th week, initial immunity: negative control group via intranasal application instillation approach awards 50 microlitre cell pyrolysis liquids after the Cavia porcellus random packet; Positive group awards 50 microlitre JSH02L, JSH02LS respectively with quadrat method, observes the Cavia porcellus situation.
In (2) the 2nd weeks, booster immunization: negative control group via intranasal application instillation approach awards 100 microlitre cell pyrolysis liquids; Positive group awards 100 microlitre JSH02L, JSH02LS with quadrat method, observes the Cavia porcellus situation.
(3) the 5th weeks, the counteracting toxic substances experiment;
1) dips in normal saline with cotton swab and clean the Cavia porcellus pudendum.
2) mix filling stomach syringe needle absorption 0.2mlHSV-2 virus with the 1mL syringe and (contain 10
6IU).
3) syringe needle is stretched into behind the intravaginal vagina arched roof viral liquid is injected, slowly withdraw from.
4) clog vagina with gelatin foam and keep viral liquid 24h.
Symptom appears in Cavia porcellus: red and swollen, blister, ulcer, incrustation.
Standards of grading:
0 is asymptomatic
0.5, only slight red and swollen;
1.0, obviously red and swollen, no vesicle;
1.5, single phlysis (≤2mm);
2.0, single big vesicle (＞2mm);
2.5, a plurality of phlysises and/or ulcer of vagina (hemorrhage);
3.0, a plurality of big vesicles;
3.5, serious swelling of external genitals;
4.0, a plurality of little/big scar merges;
4.5, back acroparalysis;
5.0, vulval ulcer, back acroparalysis
4, experimental result:
Negative control group incidence behind the expection counteracting toxic substances: all occurred just sending out infection symptoms in 3 days behind the counteracting toxic substances, about 6～7 days, Cavia porcellus skin lesion degree was the most serious, alleviated gradually, disappeared basically after the 10th day.Middle idol has the seriously ill death of Cavia porcellus.Recurrent infection occurs after 13 days after the first attack, whole scoring is bigger; Each positive group expection idol has Cavia porcellus slight red and swollen symptom to occur, does not have other manifest symptom, and whole scoring is less.Positive group and negative group significant difference, vaccine can prevent genital herpes.
Embodiment 5 recombinant herpes simplex virus I I type live vaccine are to the immunization therapy experiment of Cavia porcellus genital herpes
1, experiment material: with above-mentioned embodiment:
2, experimental technique: with behind the HSV-2 virus attack Cavia porcellus the 5th day, Cavia porcellus is divided into 5 groups at random, the via intranasal application approach gives JSH02L, each 50 microlitre of JSH02LS respectively; 14 days after the immunization therapy for the first time; Carry out the immunization therapy second time with approach and dosage, after two weeks again immunity once, treat Cavia porcellus just send out infection symptoms disappear the back with ultra violet lamp Cavia porcellus pudendum 10 minutes to bring out recurrent infection; Observe Cavia porcellus recurrence situation, calculate the skin lesion index.
3, experimental result: matched group recurrence symptom is obvious after the expection administration, and it is bigger to mark, and each positive group recurrence symptom is slight, and it is less to mark, and this vaccine has certain therapeutical effect to recurrence type genital herpes on one's body Cavia porcellus, can alleviate the recurrence symptom.
Claims (3)
1. a herpes simplex virus I I type gene recombinaton attenuated live vaccine is characterized in that knocking out the UL43～UL47 gene in its genome.
2. herpes simplex virus I I type gene recombinaton attenuated live vaccine as claimed in claim 1 is characterized in that uniting the UL43 that knocked out in its genome and duplicates or infect the dispensable gene fragment to UL47 gene and other; Said other duplicates or infect the dispensable gene fragment is that US9 is to the US12 gene.
3. the method for preparing of a herpes simplex virus I I type gene recombinaton attenuated live vaccine according to claim 1 or claim 2 is characterized in that:
Step 1: get herpes patient's BF, isolation identification goes out wild herpes simplex virus I I type;
Step 2: extract viral complete genome group design primer amplification after the amplification culture and go out the homologous flanking sequence of the virus genomic UL43 gene of herpes simplex virus I I type left side 700bp to 2000bp and the homologous flanking sequence of the virus genomic UL47 gene right side 700bp to 2000bp of herpes simplex virus I I type;
Step 3: said homologous flanking sequence of step 2 and fluorescence protein gene are cloned into the pShuttle-CMV carrier; Make fluorescence protein gene between homologous flanking sequence; Again with herpes simplex virus I I type genome cotransfection to the Vero cell, carry out homologous recombination;
Step 4: under fluorescence microscope, use a kind of or unite and use multiple fluorescence protein gene to serve as a mark to pick out recombinant virus; Through obtaining herpes simplex virus I I type gene recombinaton attenuated live vaccine behind the plaque purification.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2552341C1 (en) * | 2014-02-26 | 2015-06-10 | Федеральное государственное унитарное предприятие "Санкт-Петербургский научно-исследовательский институт вакцин и сывороток и предприятие по производству бактерийных препаратов" Федерального медико-биологического агентства (ФГУП СПбНИИВС ФМБА России) | Pharmaceutical composition for producing antiherpetic mixed vaccine and based dosage form |
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| WO2002092826A2 (en) * | 2001-05-09 | 2002-11-21 | M's Science Corporation | Composition and method for treating cancer using herpes virus |
| US20050112142A1 (en) * | 2003-04-25 | 2005-05-26 | Spaete Richard R. | Methods and compositions for treatment and prevention of HSV-2 infections and conditions |
| US7118755B2 (en) * | 1998-07-31 | 2006-10-10 | Biovex Limited | Herpes viruses for immune modulation |
| CN101926992A (en) * | 2010-08-16 | 2010-12-29 | 郑州金森生物科技工程有限公司 | Recombined attenuated live vaccine for preventing and treating I-type or II-type infection of herpes simplex virus and preparation method thereof |
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2010
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| US7118755B2 (en) * | 1998-07-31 | 2006-10-10 | Biovex Limited | Herpes viruses for immune modulation |
| WO2002092826A2 (en) * | 2001-05-09 | 2002-11-21 | M's Science Corporation | Composition and method for treating cancer using herpes virus |
| US20050112142A1 (en) * | 2003-04-25 | 2005-05-26 | Spaete Richard R. | Methods and compositions for treatment and prevention of HSV-2 infections and conditions |
| CN101926992A (en) * | 2010-08-16 | 2010-12-29 | 郑州金森生物科技工程有限公司 | Recombined attenuated live vaccine for preventing and treating I-type or II-type infection of herpes simplex virus and preparation method thereof |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2552341C1 (en) * | 2014-02-26 | 2015-06-10 | Федеральное государственное унитарное предприятие "Санкт-Петербургский научно-исследовательский институт вакцин и сывороток и предприятие по производству бактерийных препаратов" Федерального медико-биологического агентства (ФГУП СПбНИИВС ФМБА России) | Pharmaceutical composition for producing antiherpetic mixed vaccine and based dosage form |
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