SEMINAR ON CELL CULTURE BASED VACCINES

Presented byRohini dev M.Sc BIOTECHNOLOGY 3rd SEM.

CONTENTS

Introduction Types Of Vaccines Vaccine : Production Procedures -Vaccines Produced In Fertilized Chicken Eggs -Vaccine Based On Cell Or Tissue Cultures Influenza Vaccines -The Burden Of Influenza -Antigenic Shift And Antigenic Drift Cell-based Vaccine Production -Regulations And Limitations Dc-based Cancer Vaccine Recent Advancements References

Introduction
What is a Vaccine?

A vaccine(Latin: vaccacow ) is a non-pathogenic antigen that mimics a particular pathogen in order to elicit an immune response The first recorded attempts to induce immunity deliberately were performed by the Chinese and Turks in the fifteenth century English physician Edward Jenner(1798)- successful use of cowpox virus to immunize people against smallpox Louis Pasteur (1880s)- developed a vaccine against anthrax and rabies , he was honoring Jenner when he applied the term vaccination to immunization with attenuated cultures of bacteria that had no connection with cows

A. SUCCESS STORY -Complete eradication of smallpox -WHO prediction : eradication of paralytic polio throughout the world by year 2003 -Significant reduction of incidence of diseases like: Diptheria,Measles,Mumps,Pertusis,Rubella, Poliomyelitis,Tetanus B. NEED OF AN HOUR - Search for non-availabile effective vaccines for diseases like: Malaria, Tuberculosis and AIDS - Improvement in safety and efficacy of present vaccines - Low cost - Efficient delivery - Reduction of adverse side effects

Types of Vaccines

Live attenuated vaccines-Vaccines against measles,mumps, and chickenpox Inactivated vaccines- The salk vaccine for polio Subunit vaccines- For the hepatitis B virus Toxoid vaccines- Vaccines against diphtheria and tetanus DNA vaccines- Vaccines for influenza and herpes Chimeric Vaccines- Yellow fever vaccine

Vaccine : production procedures

Vaccines produced in fertilized chicken eggs
o Eggs are kept in incubator and embryos of 7-12 days old are o o o o

used Each strain is grown separately Embryo infected virus multiplication Harvest purification inactivation and production 1-2 eggs require to produce 1 dose of vaccine Entire production process lasts at least six months

Advantages:

Well established Lower cost Disadvantages: Extensive planning: long timeline for million eggs procurement Limited flexibility in case of exponentially increasing demand: -production takes too long time -eggs dont grow on demand

Potential impurities in eggs (antibiotics, other viruses) may cause sterility problems Risk of allergies against egg albumin Growth of epidemic viruses in eggs result in variants that are antigenically distinct from the original viruses Emerging endemic viruses sometimes do not grow in eggs

Vaccine Based On Cell Or Tissue Cultures

Cell culture based vaccines have major

advantages over the traditional manufacturing process Mammalian kidney cells are preferably used for these cell cultures Virus is injected into these cells, multiply before cells outer walls are removed Harvest, purification, inactivation and production Process similar to biotechnological fermentation, in which production can be done in small liter jars to huge fermenters

Vaccine Production Plant

Fermenter 4 Virus Production

Fermenter 3 Cell propagation High Cell Density

Fermenter 2 Cell propagation

Fermenter 1 Cell propagation

Cell-culture Technologies May Offer Distinct Advantages Over Egg Based Manufacturing Methods:

They eliminate the need for embryonated chicken eggs from managed,biosecure flocks They combine and automate upstream and downstream processes They reduce the potential for contamination by viable and nonviable particulates They eliminate the four- to six-month lead times for the organization of egg supplies They have faster, high-volume start-up times for production They have higher initial purity They could supplement seasonal vaccine supplies when multiple strain changes are necessary They would substantially increase global stockpiles of pandemic influenza vaccines

Influenza Vaccines
Orthomyxoviridae family (RNA) Influenza viruses are of three typesInfluenza A: infect a range of animals (humans, swine, birds, seals,horses) Influenza B: infect only humans Influenza C: infects humans and swine Pleomorphic shape,enveloped virus Influenza A subtypes (surface glycoproteins antigenicity): -hemagglutinin (H1-H15) -neuraminidase (N1-N9)

The Burden of Influenza

250,000 to 500,000 deaths globally/yr 36,000 deaths and >200,000 hospitalizations/yr in U.S. $37.5 billion in economic costs/yr in U.S. related to influenza and pneumonia Ever-present threat of pandemic influenza

H9* H5*

1998 1999 2003

H7* Influenza Pandemics H1

1915 1925 1935

1997 2003-2005 1996 2002 2003 2004

1980

H1 H2
1945 1955

H3
1977
1965 1975 1985 1995

2009 Swine Flu H1N1

2005

1918 Spanish Flu H1N1

1957 Asian Flu H2N2

1968 Hong Kong Flu H3N2

2003 Avian Flu H5N1

ANTIGENIC DRIFT

ANTIGENIC SHIFT

continued Cell lines

PER C.6 (Crucell) EBx: A stem cell line derived from chicken embryos (Sigma-Aldrich Group) VERO: A kidney cell from the African green monkey (GSK) MDCK : Madin-Darby canine kidney cells used by Chiron

Requirements

Good cell-virus interaction Broadly and highly permissive for a wide variety of flu strains Restricted growth of non-flu human pathogens that may be present in the viral seed Scalable, industrializable high yield, high volume production Cell growth in chemically defined medium (no animal-derived components)

Continued..

Novartis is a member of the European Federation of Pharmaceutical Industries and Associat (EFPIA)and of the International Federation of Pharmaceutical Manufacturers and A (IFPMA) Rino Rappuoli is global head, Vaccines Research, June 13, 2007 (CIDRAP News) The Novartis Vaccines European Union (EU) has approved Novartis's seasonal influenza vaccine, Optaflu, the first flu vaccine grown in cell culture rather than eggs The vaccine has been approved for use in all 27 EU member states including Iceland and Norway

Cell-based vaccine production: regulations and limitations

Regulations Complete removal of the cells from the final product so that the cell line does not bring any transforming agent (oncogenic transformation) into the final product No genetic material is left from the cell line in the final product that can cause tumors to be formed. All residual DNA must be removed or inactivated so it cannot give rise to tumors in animal models Limitations Not all infectious agents can be grown in culture Animal/human cell culture is expensive Yield of viruses from cultures can be low Safety precautions for culture of live agents

DC-based cancer vaccine

Dendritic cells (DC), a leukocyte population,represent unique antigen-producing cells capable of sensitizing T cells to elicit antigen specific immune response DCs are the most potent antigen-presenting cells (APS) Tumor antigens in different forms (DNA, RNA, proteins, peptides, viruses, cell lysates) become immunogenic when presented to T-lymphocytes by DCs Immunization with ex-vivo generated DC has proven feasible and permits the enhancement as well as the dampening of antigen-specific immune responses in man Several DC-based clinical trials have demonstrated potent immunological and some clinical responses

Recent

Advancements

Influenza Vaccines of the Future ScienceDaily (Nov. 18, 2010) In a review article appearing in the New England Journal of Medicine, scientists at the National Institute of Allergy and Infectious Diseases (NIAID), part of the National Institutes of Health, examine research under way to address the limitations of currently available influenza vaccines and develop more efficient and reliable strategies to make vaccines to protect against seasonal as well as pandemic influenza. New Experimental Vaccine Against Chikungunya Virus Created ScienceDaily (Aug. 14, 2011) Researchers have developed a new vaccine to protect against chikungunya virus, a mosquito-borne pathogen that produces an intensely painful and often chronic arthritic disease that has stricken millions of people in India, Southeast Asia and Africa. ScienceDaily (Aug. 15, 2011) Until now. Prof. Nir Ben-Tal of Tel Aviv University's Department of Biochemistry and Molecular Biology and his graduate student Daphna Meroz, in collaboration with Dr. Tomer Hertz of Seattle's Fred Hutchinson Cancer Research Center, have developed a unique computational method or valuable tool for identifying viral mutation strategies, developing vaccinations and anti-virals which can protect the population. Published in the journal PNAS. BioSpectrum October (2010 ) Indias leading biopharma company Bharat Biotech, launch the countrys first indigenously developed cell culture H1N1 swine flu vaccine under the brand name HNVAC. This is said to be the only flu vaccine to be manufactured in cell culture, in the developing worlda highly sterile and controlled manufacturing process, instead of eggs.

REFERENCES

Kindt TJ, Goldsby RA and Osbarne BA, Kuby Immunology, WH Freeman & Company, New York, 5th edition,2007, Pg 481-488. Ed. Clynes M, Animal Cell Culture Techniques, Springer, Berlin, Heidelberg, New York, 1998,Pg13-17. Kang SM, et al. Influenza Virus-Like Particles As Pandemic Vaccines. Current Topics in Microbiology and Immunology: Vaccines for Pandemic Influenza. Springer: Berlin, Germany, 2009. Part 3, Vol. 333: pg 269289. "Dendritic cells and immunity against cancer" by K. Palucka, H. Ueno1, J. Fay, and J. Banchereau of Baylor Institute for Immunology Research and Sammons Cancer Center, Baylor University Medical Center,Dallas, TX; and Department of Gene and Cell Medicine and Department of Medicine, Immunology Institute, Mount Sinai School ofMedicine, New York, NY,USA as published in Journal of Internal Medicine.Volume 269, Issue 1, 2010. http://www.ncbi.nlm.nih.gov/pubmed/21461683 http:// www.elsevier.com/locate/vaccine http;//www.sciencedaily.com/