病毒灭活血小板临床应用进展

1. 病毒灭活血小板临床应用进展 邱艳 北京血液中心 2013年9月5日淮安

2. 为什么病毒灭活成分血？ BacteriaThe most frequenttransfusion-transmitted infection Known pathogens Routine testing covers only a limited number New and emerging pathogens A risk that current safety measures cannot eliminate Screening limitationsGaps in current defenses exist, due to the window period and limited screening sensitivity Leukocytes Residual cells and cytokines can cause harmful post-transfusion reactions Most transfusion recipients are battling serious disease and have weakened immune systems

3. Emerging Infections Disease: continue to emerge worldwide Red – newly emerging diseases: Blue – re-emerging/resurging diseases Adapted from Morens DM. Nature. 2004;430:242–9.

4. Timeline of the WNV epidemic in the USA PUBLIC HEALTH: WNV was recognized as a threat in 1999 First identification in humans, horses and birds in three states 66 human cases reported in 10 states 9,862 human cases, WNV Ab test approved WNV activity in 47 continental states, 2,470 human cases 1999 2000 2001 2002 2003 2004 2005 199 WNV-(+) donors in 28 states First documented transmission by organ transplant and blood transfusion WNV NAT implemented under IND; transmission confirmed or suspected in >40 blood recipients BLOOD SAFETY: Experimental tests for WNV were not implemented until June 2003

5. WNV isolation from mosquitoesor vertebrates, including humans Laboratory-confirmed human or equine cases of West Nile fever Presence of antibodies to WNV in vertebrates West Nile virus in Europe • Recent outbreaks of WNV in Europe reaffirmed that mosquito-borne viral diseases may occur on a mass scale, even in temperate climates1 • Frequent detection of equine, avian and human WNV in south France (2000 and 2003) suggests that some areas may be at increased risk of infection on a seasonal, rather than sporadic, basis2 1. Hubálek Z, et al. Emerg Infect Dis. 1999;5:643–50. 2. Dauphin G, et al. Comp Immunol Microbiol Infect Dis. 2004;27:343–55.

6. Healthy blood donors can carry a range of asymptomatic herpesviridae infections *Severe symptoms most likely in immunocompromised individuals. **Moderate prevalence in Mediterranean countries, low in rest of Europe. 1. http://www.cdc.gov/ncidod/diseases/cmv.htm2. http://www.cdc.gov/ncidod/diseases/ebv.htm3. Clark DA. Int J Hematol. 2002;76:246–52.

7. Studies have shown CMV transmission by CMV-negative and leukoreduced blood products *Primary and secondary endpoints (infections day 21–100 or 0–100 after SCT) Adapted from Ljungman P. Br J Haematol. 2004;125:107–16.

8. Anti-HCV NAT HIV p24 Syphilis Anti-HIV-1 1960 1965 1970 1975 1980 1985 1990 1995 2000 2005 HBsAg Anti-HBc, Anti-HTLV-I/II Leuko-depletion Anti-HIV-2 Pathogen Inactivation Availability of safety measures against transfusion-transmitted infections Adapted from Barbara, J. Transfus Med Hemother. 2004;31(suppl 1):1–10.

9. 血液制品病毒灭活的历史

10. 血浆成分血病毒灭活的现状

11. 血小板成分血病毒灭活的现状

12. 红细胞成分血病毒灭活的现状

13. 血小板成分血病毒灭活的机理 X, G Radio WAVE LENGTH (nm) High energy Low energy 200 250 300 350 400 590 750 UVC UVB UVA VISIBLE Amotosalen INTERCEPT 3 J/cm2 Mirasol 6.2 J/mL UVC 0.4 J/cm2

14. The INTERCEPT Blood System Introduction & Mechanism

15. NH2 O O O O Nucleic acid targeting using amotosalen HCl • Psoralen targets DNA and RNA (single- and double-stranded) • Crosslinking reaction is initiated by UVA light (320-400nm) • Replication of nucleic acid of pathogens and leukocytes is stopped • Platelets, plasma and RBCs do not require nucleic acid function for therapeutic effect Amotosalen

16. Amotosalen crosslinks both single- anddouble-stranded nucleic acids Helical Regions Double-strandedDNA or RNA Single-strandedDNA or RNA

17. Amotosalen mechanism of action UVA Illumination Amotosalen (S-59) Targeting Intercalation Crosslinking Helical region of single- or double-stranded DNA or RNA Multiple crosslinks block strand separation and replication

18. The INTERCEPT Blood System for platelets:A broad spectrum of inactivation • Bacteria • Aerobic and anaerobic species • Gram-positive and Gram-negative species • Spirochetes (including Treponema pallidumand Borrelia burgdorferi) • Viruses • Enveloped and non-enveloped species • Species with either single- or double-stranded DNA or RNA • Protozoa • Including Plasmodium spp. (malaria), Trypanosoma cruzi (Chagas’ disease), and Leishmania spp. (leishmaniasis) • Residual donor leukocytes • Prevents replication and inhibits cytokine synthesis

19. Limitations of the INTERCEPT Blood System • Not effective against HAV • Highly infrequent labile component pathogen with few reported cases • Not effective against bacterial spores • Effective against vegetative state • Not effective against prions • Unique disease mechanism • Very low frequency • Preventable by other measures: livestock management

20. The INTERCEPT Blood System for Platelets Virus

21. The INTERCEPT Blood System for platelets:Viral inactivation overview • Small non-enveloped picorna viruses (e.g., poliovirus, HAV) are resistant to inactivation

22. Measurement of pathogen inactivation:Log reduction • 灭活水平计算公式: Log (Pre-treatment infectivity / Post-treatment infectivity) 或 Log (Pre-treatment infectivity) – Log (Post-treatment infectivity) • 处理后没有存活病毒,用 <1 infectious unit/volume assayed表示 • “>”表示灭活效果的最低检测限 • Log reduction was reported based on the volume assayed

23. Examples of log reduction calculations

24. INTERCEPT Blood System for platelets: Inactivation of routinely tested pathogens *Approved product claims under CE Mark. “>” refers to below limit of detection for assay.

25. INTERCEPT Blood System for platelets: Inactivation of additional enveloped viruses “>” refers to below limit of detection for assay. *Approved product claims under CE mark. 1. Pinna D, et al. Transfus Med. 2005;15:269–76. 2. Lin L, et al. Transfusion. 2005;45:580–90.

26. INTERCEPT Blood System for platelets: Inactivation of non-enveloped viruses *Approved product claims under CE mark.†This incubation period was supplementary to the standard operating protocol. 1. Sawyer L. Manuscript in preparation.

27. Implementation & Data Management

28. HOSTsystem INTERCEPT data management system (IDMS) IDMS server Registration Add amotosalen Illumination CAD incubation Storage

29. Macopharma Platelets

30. 谢谢!