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Cell culture based vaccine

The document discusses the development and application of cell culture-based vaccines, highlighting various types of animal cell substrates such as primary cell cultures, diploid cell lines, and continuous cell lines. It outlines the historical context, advantages, disadvantages, and general steps involved in vaccine production from these cell types, as well as the use of dendritic cells in personalized cancer vaccines. Additionally, it identifies top biotech companies in India and mentions the potential risks and limitations of using cell culture for vaccine production.

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CELL CULTURE BASED VACCINE By KAUSHAL KUMAR SAHU Assistant Professor (Ad Hoc) Department of Biotechnology Govt. Digvijay Autonomous P. G. College Raj-Nandgaon ( C. G. )
Introduction  Cell culture based vaccine??  Cell cultures involve growing cells in a culture dish, often with a supportive growth medium. A primary cell culture consists of cells taken directly from living tissue, and may contain multiple types of cells such as fibroblasts, epithelial, and endothelial cells.  In the United States, 10 different vaccines for chicken pox, hepatitis A, polio, rabies, and rubella are cultured on aborted tissue from two fetal cell lines known as WI-38 and MRC-5. These vaccines are chicken pox, hep-A, hep-A, hep-A/hep-B, polio, rabies, rubella, measles/rubella, mumps/rubella, and MMR II (measles/mumps/rubella).
History  PCCs of monkey kidney cells have been used for the production of inactivated and oral poliomyelitis vaccines since the 1950s.  In 1954, an experimental adenovirus vaccine was being developed, and human tumour cells (HeLa) were rejected as the cell substrate in favour of ”normal” cells.  The first requirements for cell substrates were published by WHO in 1959 for the production of inactivated poliomyelitis vaccine in PCCs derived from the kidneys of clinically healthy monkeys.  In the 1960s, human diploid cells (HDCs) were developed and proposed as an alternative to primary monkey kidney cell cultures for polio virus vaccine production as well as for other viral vaccines.  WHO Requirements for Continuous Cell Lines used for Biologicals Production were published in 1987.  During the 1990s, and on into the 2000s, a variety of CCLs were explored as cell substrates for biological products.
Types of animal cell substrates 1. Primary Cell Cultures (PCCs) o Major successes in the control of viral diseases, such as poliomyelitis, measles, mumps and rubella, were made possible through the wide use of vaccines prepared in PCCs, including those from chicken embryos and the kidneys of monkeys, dogs, rabbits and hamsters. Advantages: (a) Easy to prepare using simple media and bovine serum; (b) Broad sensitivity to various viruses, some of which are cytopathic. Disadvantages: (a) Contamination by infectious agents is a higher risk than with DCLs and CCLs; (b) They cannot be tested as extensively as DCLs or CCLs.
Types of animal cell substrates 2. Diploid Cell Lines (DCLs) The practicality of using human DCLs for the production of viral vaccines was demonstrated in the 1960s. DCLs of human (e.g., WI-38, MRC-5) and monkey ( i.e., FRhL2 ) origin. Advantages (a) They can be well characterized and standardized; (c) Unlike the CCLs and SCLs , DCLs are not tumourigenic and therefore do not raise the potential safety issues associated with CCLs and SCLs. Disadvantage (a) In general, they have more fastidious nutritional requirements than other cell substrates; (b) They may be difficult to adapt to serum-free growth; (c) They are more difficult than CCLs to transfect and engineer, and require immortalization before they can be engineered;
Types of animal cell substrates 3.Continuous cell lines (CCLs) CCLs have the potential for an apparently indefinite in vitro life span and have been derived by the following methods: a) Serial subcultivation of a PCC of a human or animal tumour (e.g., HeLa cells); (b) Transformation of a normal cell having a finite life span with an oncogenic virus or viral sequence (e.g., B lymphocytes transformed by EBV or transfected with viral sequences such as in PER.C6); (c) Serial subcultivation of a primary-cell population derived from normal tissue that generates a dominant cell population having an apparently indefinite life span, often described as spontaneous transformation (e.g., Vero, BHK-21, CHO, MDCK, Hi5); (d) Fusion between a myeloma cell and an antibody-producing B lymphocyte to produce a hybridoma cell line.
Advantages: (a) Characterized extensively and their culture conditions standardized; (b) Grow more easily than DCLs using standard media, (d) Most can be adapted to grow in serum-free medium; (e) Can be grown on micro-carriers for large-scale production in bioreactors; (f) some can be adapted to grow in suspension cultures for large-scale production in bioreactors. Disadvantages (a) CCLs may express endogenous viruses, and some are tumourigenic in immuno-suppressed animal models; (b) Theoretical risks identified by the 1986 Study Group (e.g., nucleic acids, transforming proteins, and viruses) need to be taken into account.
GENERAL STEPS OF VACCINE PRODUCTION FROM ANIMAL CELLS
VACCINES FROM CELL CULTURE  MMR Vaccine  Polio vaccine  Chickenpox (Varicella) vaccine  Rabies vaccine  Recombinant protein vaccines  Influenza vaccine  Dendritic Cell-Based Vaccination in Solid Cancer
MMR Vaccine Disease Immunized Virus Strain Propagation cell Growth Medium Measles Enders' attenuated Edmonston strain chick embryo cell culture Medium 199 Mumps Jeryl Lynn(B level) strain chick embryo cell culture Medium 199 Rubella Wistar RA 27/3 strain of live attenuated rubella virus WI-38 human diploid lung fibroblasts MEM (solution containing buffered salts, fetal bovine serum, human serum albumin and neomycin, etc.)
Polio , Chickenpox and Rabies vaccine Disease Immunized Virus Strain Propagation cell Growth Medium Poliomyelitis or polio Type 1 (Mahoney), Type 2 (MEF-1), Type 3 (Saukett) vero cells, a continuous line of monkey kidney cells Eagle MEM modified medium, M-199 Chickenpox Oka/Merck strain (varicella virus) Human diploid cell cultures (WI-38, MRC-5) MEM Rabies Attenuated Pitman-Moore L503 strain Attenuated Wistar strain Human diploid cell chicken embryo Vero cell Eagle MEM modified medium, M-199
iNfluenza vaccine
(VLPS)-based Influenza Vaccines
DC-based cancer vaccine • Dendritic cells (DC), a leukocyte population, represent unique antigen-producing cells capable of sensitizing T cells to both new and recall antigens. • 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
DC loading and activation  Ex vivo generated dendritic cells(DC) can be loaded with antigens and re-infuse to the patients, or they can be used for ex vivo expansion of anti-tumor lymphocytes.
Personalized Cancer Vaccine More than 150 DC-based clinical studies for the treatment of solid or hematological malignancies have been reported in 2008 including;  Melanoma (〉 40 published clinical studies)  Prostrate cancer (20)  Renal cell carcinoma (16)  Breast cancer (12)  Multiple myeloma (9)  Leukemia (9)  Colorectal cancer (9)  Glioma (9) Nencioni, A. et.al. Critical Reviews in Oncology/Hematology 65(2008), 191-199
Advantages  Eliminate the need for embryonated chicken eggs from managed, biosecure flocks.  Combine and automate upstream and downstream processes.  Reduce the potential for contamination by viable and nonviable particulates.  Eliminate the four- to six-month lead times for the organization of egg supplies.  They have faster, high-volume start-up times for production.  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.
Potential risks  Viruses and other transmissible agents  Cellular DNA  Cellular RNA  Limitations of Vaccine Production using cell culture  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
The top biotech companies in India  Biocon : Bangalore, Karnataka  Serum Institute of India :Pune, India  Panacea Biotech : Mumbai  Shantha Biotech : Hyderabad  Indian Immunologicals : Hyderabad, Ooty and Rajkot  Bharat biotech : Hyderabad  Syngene International : Bangalore
REFERENCES  Springer.com  WHO official website  www.biopharminternational.com