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Pharmacy & Pharmacology International Journal

Review Article Volume 8 Issue 4

3Rs expression in quality control paradigms of human vaccines 

Syeda Shazia Adeel,1 Tanveer Ahmed Khan,1 Kanwal Batool,1 Baseer Ahmad Khan2

1National Institute of Health, Pakistan
2Institute of Biochemistry and Biotechnology, University of Veterinary & Animal Sciences, Pakistan

Correspondence: Tanveer Ahmed Khan, National Institute of Health, Islamabad, Pakistan

Received: May 23, 2020 | Published: July 13, 2020

Citation: Adeel SS, Khan TA, Batool K, et al. 3Rs expression in quality control paradigms of human vaccines. Pharm Pharmacol Int J. 2020;8(4):201-207. DOI: 10.15406/ppij.2020.08.00297

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Abstract

Communicable diseases remain the leading cause of mortality worldwide in past. Children and adolescent were the most affected individuals. However, the development of vaccine played a vital role in decreasing mortality and increasing life expectancy. Currently most of vaccines are based on utilizing animal pathogens and use of animals. This use of animal in quality testing of vaccines is inevitable. Over the past century, concern about animals’ interests was limited to ensure that animals be treated humanely and not subjected to unnecessary sacrifice and sufferings. However, with the rise of 3Rs concept the global scenario for use of animals is being changed a little. The current review addresses the refinement, reduction and replacement aspects of animal use in vaccine testing. It also dialogues about challenges to implement 3Rs and gadget this key concept for effective quality testing.The acceptability and implementation of 3Rs concept is based on good manufacturing practices, good in-process quality control and validated procedures and processes. It is easier for new vaccines to adopt this concept. However, several difficulties are still experienced in theimplementation of 3Rs principles for vaccine potency assays. Thus, insistent exploration is obligatory by bothhealth industries and regulatory agencies for the implementation and validation of robust 3Rs approaches around the globe.

Keywords: vaccination, quality control, 3Rs, potency testing

Introduction

Communicable diseases remain the leading cause of mortality worldwide in past. Children and adolescent were the most affected individuals. However, the development of vaccine played a vital role in decreasing mortality and increasing life expectancy. Vaccines prevented 178 million cases from 1888 to 1924 in United States only.1 Similarly, 300 million people died of smallpox in the 20th century but no one dies today due to vaccination.2 Worldwide life expectancy has also been increased.3 Vaccines are being developed using the principles set by Louis Pasteur since more than 100 years.4 In last few decades, several technologies have been industrialized to develop vaccines.5

The current review addresses the refinement, reduction and replacement aspects of animal use in vaccine testing.It also dialogues about challenges to implement 3Rs and gadget this key concept for effective quality testing.

The vaccines spectrum

Vaccines have saved more lives than any other medicine or medical product around the globe. It has been proved very effective tool of immunization in modern medicine.6 There are many types of vaccines (Table 1) which are categorized by the antigen used in their preparation.7

Class          

Type

Nature

Example

I

Live attenuated vaccines

contain whole bacteria or viruses which have been “weakened” so that they create a protective
immune response but do not cause disease in healthy people

Rotavirus vaccine, MMR vaccine, Nasal flu vaccine, Shingles vaccine, Chickenpox vaccine, BCG vaccine against TB, Yellow fever vaccine, Oral typhoid vaccine (not the injected vaccine)                                                                                                                                                                                                                                     

II

Inactivated vaccines

contain whole bacteria or viruses which have been killed, or small parts
of bacteria or viruses, such as proteins or sugars, which cannot cause disease

IIA

Whole killed’ vaccines

contain whole killed viruses

Inactivated polio vaccine or IPV, Some inactivated flu vaccines which are described as ‘split virion’, Hepatitis A vaccine, Rabies vaccine, Japanese encephalitis vaccine

IIB

Subunit vaccines (sometimes called ‘acellular’)                                                                                        

do not contain any whole bacteria or viruses at all. (‘Acellular’ means ‘not containing any whole cells’.) Instead contain polysaccharides
(sugars) or proteins from the surface of bacteria or viruses. These polysaccharides or proteins are the parts that our immune system recognizes as ‘foreign’, and they are referred to as antigens

IIB-1

Toxoid vaccines

Some bacteria release toxins (poisonous proteins) when they attack the body. The immune system recognizes these toxins in the same way that it recognizes polysaccharides or proteins on the surface of the bacteria. Some vaccines are made with inactivated versions of these toxins. They are called ‘toxoids’
because they look like toxins but are not poisonous                                                                                                                                                                                                                                              

Diphtheria vaccine, Tetanus vaccine, Pertussis (whooping cough) vaccine

IIB-2

Conjugate vaccines

In most conjugate vaccines, the polysaccharide is attached to diphtheria or tetanus toxoid protein. The immune system recognizes
these proteins very easily and this helps to generate a stronger immune response to the polysaccharide

Hib vaccine, MenC vaccine, PCV

IIB-3

Recombinant vaccines

made using bacterial or yeast cells to manufacture the vaccine. A small piece of DNA is taken from the virus or bacterium against which we want to protect. This is inserted into other cells to make them produce large quantities
of active ingredient for the vaccine (usually just a single protein or sugar)

Hepatitis B vaccine, MenB vaccine, inactivated flu vaccines described as ‘surface antigen’, PPV, Injected typhoid vaccine (a polysaccharide vaccine)

Table 1 Types of vaccines with examples

Quality control testing for vaccines

Biological products are different in comparison to other pharmaceutical products; same is the case with vaccines. They are being derived from activemicroorganisms. However, their composition is complicated to be explained in perspective of chemical of physical means. Moreover, the intrinsic diversity of microorganisms and the potential for contamination of materials use in vaccine production requires special attention for quality control of vaccines.8 Generalized tests of vaccines for human use include pH, Adjuvant, Aluminium, Calcium, Free Formaldehyde, Phenol, Water, Extractable volume and Bacterial endotoxins.9 Typically, individual vaccine may contain the following tests;9

  1. Residual pertussis toxin for vaccines (comprising acellular pertussis factor)
  2. Residual infectious virus
  3. Sterility test (for live vaccines)
  4. Pyrogens/Bacterial endotoxins
  5. Total protein content (where relevant)
  6. Free saccharide (for conjugate vaccines)
  7. Potency test
  8. Distribution of molecular size/ molecular weight (for polysaccharide vaccines)
  9. Ovalbumin content (where a vaccine is produced in eggs)
  10. Bovine serum albumin (where a vaccine is produced in cell cultures)
  11. Host cell and vector DNA
  12. Host cell protein
  13. Residual reagents
  14. Vesicle size (virosomal vaccines) 

Use of animals in quality control

Research on infections and infestations in veterinarycontributed a significant part in the advancement of vaccines for human use over past century. Currently most of vaccines are based on utilizing animal pathogens.10 Likewise, use of animals in quality testing of vaccines has become an important tool. Animals are used in vaccine development and testing to evaluatesafety of vaccine; defense against an infection or illness; decreasein clinical signs &symptoms;death of pathogen; commencement and extent of immunity; category of immune response; routes of vaccine administration; and evaluation of particular immune compartments.11 Rats, mice12 or guinea pigs are used in potency tests for vaccines. There is a huge diversity of potency tests for vaccines especially those labelled as bacterial. Code of Federal Regulations specified few standards for vaccines like cholera, typhoid, pertussis, anthrax and BCG vaccines while others like tetanus, plague and diphtheria vaccines follow minimum requirements. On contrary, acellular pertussis, polysaccharide conjugate and live oral typhoid are analyzed according to adapted standards.13 With the diversity of potency testing, large numbers of animals are used in testing procedures.There are no exact figures available for the use of laboratory animal in quality testing of vaccines. However, there is an estimate of 10% laboratory animals being used in biomedical research and testing. It includes more than one million rodents and guinea pigs only in European countries.14 Various approaches were adopted to reduce number of animals in animal testing. However, these approaches were based on a change in experimental design, a change based on statistical review and changes resulting from harmonization of test requirements.15 These approaches accompanied the concept of 3Rs. 

3Rs concept in vaccine quality control

WilliamRussel and Rex Burch formulated the principles for humane technique firstly in 1959.16 A milestone was achieved as 3Rs concept in quality control of vaccines in an international forum in Londonin the year 1985. This concept of 3Rs can be translated in the following context:

  1. Replacement means implementing methods which avoid or replace the use of animals
  2. Reduction means changing the test design in order to minimize the number of animals per experiment
  3. Refinement means moving to methods that minimize suffering and improve animal welfare (e.g. replacing challenge tests by immunogenicity assays)

Europe converted this concept into a legal requirement and documented it in European medicines agency guidelines 1997.17 It was followed by a directive published in 2001 for both veterinary and medicinal products.18 However, the main ordinance was issued in June, 2010 on safe use of animals for scientific research and testing.19 

Prominence of 3Rs in vaccinology

The development of robust assay in vaccine demands the mechanism for induction of safe immune response and action of pathogen or pathogenic entity in causing disease.Furthermore,this assay development necessitates the insight of virulence factors that exert theirpathogenic effects. Development of assays considering these principles will result in complete replacements of animal models. However, lack of scientific knowledge at presentlimits the development of such mechanism based assays.Currently, the concept of vaccine quality control is being shift from classical model to consistency approach.20 Thus, the foremost attention should endure in monitoring of consistency rather than to establish the factual efficacy of a vaccine.21 Four tiers are being used to implement and justify the concept of 3Rs in quality control of vaccines (Figure 1). However, Figure 2 shows few examples of 3Rs for human vaccines potency testing.

Figure 1 Four tiers of 3Rs.

Figure 2 Matrix for 3Rs expression of potency testing (human vaccines).

Challenges to implementing 3Rs

World health organisation directed to national quality control laboratories to apply the concept of 3Rs. However, there are two main challenges to implementing 3Rs. First is scientific and second is regulatory. Scientific challenge involves the inherent variability of in vivo assays, validation issues of in vivo assays as per ICH guidelines50 and the attributes of product quality. On the other hand, the regulatory challenge implicates the lack of harmonization in regulatory standards around the globe,51 complexity of regulatory changes and discretion of health authorities to consent deviation from established guidelines. Therefore, a one-to-one comparison is often challenging and not necessarily justified.52

Conclusion

The acceptability of 3Rs concept is based on good manufacturing practices, good in-process quality control and validated procedures and processes. However, it is easier for new vaccines to adopt this concept. Significantdevelopments have been made in the improvement, maintenance, and upgradation of in-vitro potency assays likeELISA, Gel electrophoresis, Cell culture etc. which minimize the animal use and suffering.However, several difficulties are still experienced in theimplementation of 3Rs principles for vaccine potency assays. Thus, insistent exploration is obligatory by bothhealth industries and regulatory agencies for the implementation and validation of robust 3Rs approaches around the globe.

Acknowledgments

None.

Conflicts of interest

Authors declare no conflict of interest.

References

  1. Van Panhuis WG, John Grefenstette, Su Yon Jung, et al. Contagious diseases in the United States from 1888 to the present. The New England Journal of Medicine. 2013;369(22):2152–2158.
  2. Oldstone MB. Viruses, Plagues, and history: past, present and future. Oxford University Press; 2009.
  3. Wang H, Dwyer-Lindgren L, Lofgren K, et al. Age-specific and sex-specific mortality in 187 countries, 1970–2010: a systematic analysis for the Global Burden of Disease Study 2010. The Lancet. 2012;380(9859):2071–2094.
  4. Pasteur L. De l’attenuation du virus du cholera des poules. CR Acad Sci. 1880;91:673–680.
  5. Bramwell VW, Perrie Y. The rational design of vaccines. Drug Discovery Today. 2005;10(22):1527–1534.
  6. Kumar S, Mahendra PS, Vijay KB, et al. Quality control of vaccines-A journey from classical approach to 3Rs. Microbiology: Current Research. 2018;2(3).
  7. Types of Vaccines. Vaccines Knowledge Project; 2019.
  8. Quality Control of Vaccines. WHO; 2017.
  9. Keital S. Guide for the elaboration and use of monographs on vaccines and immunosera for human use; 2019.
  10. Gerdts V, Griebel PJ, Babiuk LA, et al. Use of animal models in the development of human vaccines. Future Microbiol. 2007;2(6):667–675.
  11. Gerdts V, Wilson HL, Meurens F, et al. Large animal models for vaccine development and testing. ILAR Journal. 2015;56(1):53–62.
  12. Bruckner L, Palatini M, Ackermann M, et al. Reduction of the number of mice used for potency testing of human and animal rabies vaccines. Experientia. 1988;44(10):853–857.
  13. Habig WH. Potency testing of bacterial vaccines for human use. Veterinary Microbiology. 1993;37(3-4):343–351.
  14. Hendriksen CM, et al. Alternatives to animal testing in the quality control of immunobiologicals: current status and future prospects. ATLA. 1994;22(6):420–434.
  15. Kulpa-Eddy J, Srinivas G. Approaches to reducing animal numbers in vaccine potency testing. Procedia in Vaccinology. 2011;5:227–231.
  16. Russell WMS, Burch RL. The principles of humane experimental technique. Methuen; 1959.
  17. Replacement of animal studies by in vitro models. EEC; 1997.
  18. Directive 2001/82/EC of the European Parliament and of the Council of 6 November 2001 on the Community code relating to veterinary medicinal products. Official Journal L. 2004;311(28/11):1–66.
  19. Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. Off J Eur Union. 2010;53(L 276):33–79.
  20. Hendriksen C, Keith Redhead, Jo McKelvie, et al. The consistency approach for the quality control of vaccines. Biologicals. 2008;36(1):73–77.
  21. Hendriksen C, Spieser JM, Akkermans A, et al. Validation of alternative methods for the potency testing of vaccines. Altern Lab Anim. 1998;26(6):747–761.
  22. Xing D, Yuen C, Rigsby P, et al. Evaluation of an in vitro assay system as a potential alternative to current histamine sensitization test for acellular pertussis vaccines. Biologicals. 2012;40(6):456–465.
  23. Vaccine Acellular Component, Absorbed, in European Pharmacopoeia. 2008, Strasbourg, France: European Department for the Quality of Medicines within the Council of Europe; 2008.
  24. Hendriksen CF. Humane endpoints in vaccine potency testing. Procedia in Vaccinology. 2011;5:221–226.
  25. Xing D, Markey K, Gaines Das R, et al. Whole-cell pertussis vaccine potency assays: the Kendrick test and alternative assays. Expert review of vaccines. 2014;13(10):1175–1182.
  26. Arciniega JL, Domínguez-Castillo RI. Development and validation of serological methods for human vaccine potency testing: case study of an anthrax vaccine. Procedia in Vaccinology. 2011;5:213–220.
  27. Chang HS, Sack DA. Development of a novel in vitro assay (ALS assay) for evaluation of vaccine-induced antibody secretion from circulating mucosal lymphocytes. Clin Diagn Lab Immunol. 2001;8(3):482–488.
  28. Winsnes R, et al. Collaborative study for validation of serological methods for potency testing of diphtheria toxoid vaccines. Pharmeuropa Bio. 2004;2003(2):35–68.
  29. Winsnes R, et al. Collaborative study for the validation of serological methods for potency testing of diphtheria toxoid vaccines-part 2. Pharmeuropa Bio. 2006;2006(1):73–88.
  30. Assay of Diphtheria vaccine (adsorbed). European Pharmacopoeia: Strasbourg, France; 2008.
  31. Recommendations for diphtheria, tetanus, pertussis and combined vaccines (Amendments 2003). WHO Expert Committee on Biological Standardization. Fifty-fourth report. Geneva: World Health Organization; 2005.
  32. Assay of diphtheria vaccine (adsorbed). European Pharmacopoeia: Strasbourg, France; 2008.
  33. Kim HW, Kyung-Hyo Kim, JiHye Kim, et al. A high throughput serum bactericidal assay for antibodies to Haemophilus influenzae type b. BMC infectious diseases. 2016;16(1):473.
  34. Biological assays: Assay of hepatitis B vaccine (rDNA). Strasbourg, France: European Department for the Quality of Medicines & HealthCare; 2011.
  35. Immunochemical methods, in European Pharmacopoeia. Strasbourg, France: European Department for the Quality of Medicines & HealthCare; 2008.
  36. Shank-Retzlaff M, Wang F, Morley T, et al. Correlation between mouse potency and in vitro relative potency for human papillomavirus type 16 virus-like particles and Gardasil® vaccine samples. Human Vaccines. 2005;1(5):191–197.
  37. Kim BC, Kim DK, Kim HJ, et al. A collaborative study of an alternative in vitro potency assay for the Japanese encephalitis vaccine. Virus Res. 2016;223:190–196.
  38. Schalk J, de Vries C, Jongen P. Potency estimation of measles, mumps and rubella trivalent vaccines with quantitative PCR infectivity assay. Biologicals. 2005;33(2):71–79.
  39. Requirements for Oral Poliomyelitis Vaccine. WHO Expert Committee on Biological Standardization. Fifty-first report. Geneva: World Health Organization; 2002.
  40. WHO Expert Committee on Biological Standardization: forty-eighth report. World Health Organization; 1999.
  41. Dragunsky E, Nomura T, Karpinski K, et al. Transgenic mice as an alternative to monkeys for neurovirulence testing of live oral poliovirus vaccine: validation by a WHO collaborative study. Bulletin of the World Health Organization. 2003;81(4):251–260.
  42. CFR, Title 9 (Animals and Animal Products), Subchapter A (Animal Welfare). Office of the Federal Register Washington DC; 1985.
  43. Lin G, Chen S, Zhao H, et al. A time-resolved fluoroimmunoassay to assay the rabies virus glycoprotein: application for estimation of human rabies vaccine potency. Scientific reports. 2017;7(1):1–9.
  44. Ranheim T, Mathis PK, Joelsson DB, et al. Development and application of a quantitative RT-PCR potency assay for a pentavalent rotavirus vaccine (RotaTeq®). Journal of virological methods. 2006;131(2):193–201.
  45. Wang DY, et al. Evaluation and validation of potency testing method for live rubella virus vaccine. Journal of Food and Drug Analysis. 2001;9(4):183–190.
  46. Leparc-Goffart I, Poirier B, El Zaouk A, et al. New generation of cell culture assay for smallpox vaccine potency. J Clin Microbiol. 2003;41(8):3687–3689.
  47. Assay of Tetanus vaccine (adsorbed). European Pharmacopoeia; 2003.
  48. Hendriksen C, Cussler K, Halder M. ECVAM's role in the implementation of the Three Rs concept in the field of biologicals. Alternatives to Laboratory Animals. 2002;30(2_suppl):41–46.
  49. NICEATM-ICCVAM, Alternative Methods for Vaccine Testing, Serology Test for Batch Potency Testing of Human Tetanus Vaccines; 2012.
  50. ICH Harmonized Tripartite Guideline Q2 (R1): Validation of analytical procedures; 2005.
  51. Casey W, Brown K, Jones B, et al. Improving animal welfare and reducing animal use for human vaccine potency testing: state of the science and future directions. Procedia in Vaccinology. 2011;5:33–46.
  52. Uhlrich S, Coppens E, Moysan F, et al. 3Rs in Quality Control of Human Vaccines: Opportunities and Barriers. Alternatives to Animal Testing. Springer; 2019. 76–82 p.
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