DK2939025T3 - Cellemedieret immunresponsassay - Google Patents
Cellemedieret immunresponsassay Download PDFInfo
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- DK2939025T3 DK2939025T3 DK13869475.7T DK13869475T DK2939025T3 DK 2939025 T3 DK2939025 T3 DK 2939025T3 DK 13869475 T DK13869475 T DK 13869475T DK 2939025 T3 DK2939025 T3 DK 2939025T3
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
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- G01N33/5091—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing the pathological state of an organism
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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Claims (19)
- CELLEMEDIERET IMMUNRESPONSASSAY1. Fremgangsmåde til måling af cellemedieret immunresponsaktivitet, hvilken fremgangsmåde omfatter: (a) tilvejebringelse af en inkubationssammensætning ved at bringe en prøve omfattende immunceller, der er i stand til at producere immun-effektormolekyler efter stimulering med et peptidantigen, i kontakt med mindst ét peptidantigen med en længde valgt fra 5 til 50 aminosyrer og med mindst ét ikke-reducerende sukker, der er i stand til at forøge interferon-gamma-responset for immunceller, som reagerer på antigenet, og (b) detektering af tilstedeværelsen eller niveauet af mindst ét immun-effektormolekyle, hvor detektering af tilstedeværelsen eller niveauet af mindst ét immun-effektormolekyle omfatter detektering af tilstedeværelsen eller niveauet af interferongamma (INF-gamma).
- 2. Fremgangsmåde ifølge krav 1, hvor det ikke-reducerende sukker er et ikke-reducerende disaccharid, fortrinsvis valgt blandt trehalose og saccharose.
- 3. Fremgangsmåde ifølge krav 1 eller 2, hvor koncentrationen af det ikke-reducerende sukker i inkubationssammensætningen er mindst 1,5 mg/ml, fortrinsvis mindst 2 mg/ml.
- 4. Fremgangsmåde ifølge et eller flere af kravene 1 til 3, hvor prøven i trin (a) bringes i kontakt med en sammensætning, som omfatter antigenet og det ikke-reducerende sukker, og hvor sammensætningen fortrinsvis er indeholdt i en prøveopsamlingsbeholder.
- 5. Fremgangsmåde ifølge krav 4, hvor sammensætningen yderligere omfatter et antikoagulerende middel, fortrinsvis heparin, og hvor prøven er en fuldblodsprøve.
- 6. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 5, hvor prøven har én eller flere af følgende egenskaber: i) prøven blev indhentet fra et humant individ ii) prøven blev indhentet fra et humant individ, som er immunsvækket eller immundeficient iii) prøven omfatter immunceller udvalgt fra gruppen bestående af NK-celler, T-celler, B-celler, dendritiske celler, makrofager og monocytter; og/eller iv) prøven er fuldblod.
- 7. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 6, hvor antigenet har én eller flere af følgende egenskaber: i) et eller flere peptider anvendes som antigen, hvor det ene eller flere peptider har en længde valgt fra 7 til 50 aminosyrer ii) antigenet tilvejebringes af et eller flere syntetiske peptider iii) antigenet tilvejebringes af mindst to sæt peptider, idet et første sæt omfatter mindst et peptid med fra ca. 7 til 14 aminosyrerester i længden, og idet et andet sæt omfatter mindst et peptid med fra 15 aminosyrerester eller flere, hvor peptiderne omfatter hele eller en del af et proteinantigen iv) antigenet tilvejebringes af et eller flere peptider, der genkendes af en CD8+ cytotoksisk T-celle; og/eller v) antigenet tilvejebringes af et eller flere peptider, der genkendes af en CD8+ cytotoksisk T-celle, og hvor det ene eller flere peptider har en længde på mindre end 15 aminosyrer, fortrinsvis med en længde valgt fra 7-14 aminosyrer.
- 8. Fremgangsmåde ifølge et eller flere af kravene 1 til 7, hvor der i trin (a) anvendes to eller flere forskellige antigener, og/eller hvor der i trin (b) detekteres to eller flere forskellige effektormolekyler.
- 9. Fremgangsmåde ifølge et eller flere af kravene 1 til 8, hvor antigenet er et sygdomsspecifikt antigen, især et patogenspecifikt antigen.
- 10. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 9, hvor antigenet har én eller flere af følgende egenskaber: i) antigenet er associeret med eller er repræsentativt for en sygdom eller tilstand, for hvilken det cellemedierede immunrespons skal testes ii) antigenet stammer fra og er således krydsreaktivt med et antigen fra et patogen forbundet med en sygdomstilstand eller er et tumorassocieret antigen forbundet med en cancer iii) antigenet er et patogenspecifikt antigen, og hvor patogenet er en bakterie, et virus, en parasit, gær eller svamp iv) antigenet er et patogenspecifikt antigen, nemlig et bakteriespecifikt antigen, og hvor bakterien er valgt fra grampositive og gramnegative mikroorganismer, især mycobakteriearter, såsom mycobacterium tuberculosis, stafylokokarter, streptokokarter, escherichia coli, salmonellaarter, clostridium-arter, shigella-arter, proteus-arter, bacillus-arter, hemophilus-arter og borrelia-arter v) antigenet er et patogenspecifikt antigen, nemlig et virusspecifikt antigen, og hvor virusset er valgt blandt hepatitisvirus, såsom hepatitis B-virus og Hepatitis C-virus, herpesvirus, CMV-virus og humant immundefektvirus (HIV), og/eller vi) antigenet er fra eller er specifikt for et virus, fortrinsvis cytomegalovirus (CMV), og tilvejebringes af et eller flere peptider med en længde på 7 til 14 aminosyrerester, 7 til 13 aminosyrerester eller 8 til 12 aminosyrerester.
- 11. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 10 til overvågning eller bestemmelse af tilstedeværelsen, fraværet, niveauet eller stadiet af en sygdom eller tilstand valgt fra gruppen bestående af en infektion med et patogent middel, en autoimmun sygdom, en cancer, en inflammatorisk tilstand, eksponering for et toksisk middel, respons på et terapeutisk middel, en immundefekt og en immunosuppression.
- 12. Fremgangsmåde ifølge et hvilket som helst af kravene 1 til 11, omfattende: (a) tilvejebringelse af en inkubationssammensætning ved at bringe en fuldblodsprøve opnået fra et humant individ i kontakt med en sammensætning omfattende mindst et peptidantigen med en længde valgt fra 5 til 50 aminosyrer og mindst et ikke-reducerende disaccharid, fortrinsvis trehalose eller saccharose, og inkubation af inkubationssammensætningen i mindst 2 timer; og (b) måling af tilstedeværelsen eller niveauet af IFN-gamma frigivet på grund af stimuleringen med antigenet; hvor tilstedeværelsen eller mængden af detekteret IFN-gamma indikerer niveauet af cellemedieret immunrespons hos det humane individ.
- 13. Fremgangsmåde ifølge et eller flere af kravene 1 til 12, hvor det ikke-reducerende sukker er valgt blandt trehalose, mannitol, saccharose og raffinose, og hvor det ikke-reducerende sukker fortrinsvis er trehalose.
- 14. Sammensætning til induktion af et cellemedieret immunrespons i en prøve, omfattende: a) mindst et peptidantigen med en længde valgt fra 5 til 50 aminosyrer, hvor antigenet er et sygdomsspecifikt antigen b) mindst ét ikke-reducerende sukker, der er i stand til at forøge interferon-gamma-responset for immunceller, som reagerer på antigenet c) mindst ét antikoagulerende middel, hvor antikoagulanten er heparin.
- 15. Sammensætning ifølge krav 14, der har en eller flere af de følgende egenskaber: i) det ikke-reducerende sukker er et ikke-reducerende disaccharid, fortrinsvis valgt blandt trehalose og saccharose ii) det ikke-reducerende sukker er valgt blandt trehalose, mannitol, saccharose og raffinose iii) det ikke-reducerende sukker er trehalose iv) antigenet har en eller flere af egenskaberne som defineret i et eller flere af kravene 7 til 10 v) antigenet tilvejebringes af et eller flere peptider, fortrinsvis syntetiske peptider, der haren længde på mindre end 15 aminosyrer, fortrinsvis med en længde valgt fra 7-14 aminosyrer; og/eller vi) sammensætningen er en spraytørret sammensætning.
- 16. Prøveopsamlingsbeholder omfattende sammensætningen ifølge krav 14 eller 15.
- 17. Kit til måling af cellemedieret immunresponsaktivitet hos et individ, der omfatter mindst ét peptidantigen, der har en længde valgt fra 5 til 50 aminosyrer, mindst et ikke- reducerende sukker, der er i stand til at forøge interferon-gamma-responset for immunceller, som reagerer på antigenet, heparin, mindst en prøveopsamlingsbeholder og mindst ét detekteringsmiddel til interferon-gamma.
- 18. Kit ifølge krav 17, hvor prøveopsamlingsbeholderen har prøveopsamlingsbeholderens egenskaber ifølge krav 16.
- 19. Anvendelse af et ikke-reducerende sukker i et immunologisk assay til måling af cellemedieret responsaktivitet til et peptidantigen med en længde valgt fra 5 til 50 aminosyrer, fortrinsvis fra 7 til 50 aminosyrer, hvor tilsætningen af det ikke-reducerende sukker under inkubation af prøven med antigenet øger frigivelsen af interferon-gamma fra immunceller, der reagerer på peptidantigenet, der testes i analysen, og hvor det ikke-reducerende sukker fortrinsvis har én eller flere af følgende egenskaber: i) det ikke-reducerende sukker er et disaccharid ii) det ikke-reducerende sukker er valgt blandt trehalose og saccharose iii) det ikke-reducerende sukker er valgt blandt trehalose, mannitol, saccharose og raffinose iv) det ikke-reducerende sukker er trehalose v) det ikke-reducerende sukker anvendes i en koncentration på mindst 1 mg/ml, fortrinsvis 2 mg/ml i inkubationssammensætningen omfattende prøven, der skal testes, og antigenet vi) det ikke-reducerende sukker tilvejebringes i form af en sammensætning, der yderligere omfatter antigenet, som skal testes, og eventuelt et antikoaguleringsmiddel.
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