Vaccines Test 1

• 1. WHAT:
• A.protein that binds specifically to a particular substance- (substance is antigen)
• B. each= unique structure so that it can bind to its particular antigen
• C. all similar structure in immunoglobin category
• 2. ORIGIN:
• A. produced by differentiated B-cells           (plasma cells) in response to infection or immunization
• Purpose:
• 3.  bind to antigen and neutralize pathogens or prepare them for uptake and destruction by phagocytes.

• performed by antibodies of macrophage
• 1. neutralization
• 2. opsonization
• 3. complement activation

• 1. there is a toxin. and cell has a receptor on membrane for toxin.
• 2. antibody then attaches to toxin which detaches to membrane
• 3. antibody-toxin complex ingested by macrophage
• ANTIBODY PREVENTS bacterial ADHERENCE

• 1. bacteria is in extracellular space
• 2. antibody identifies specific bacteria and antigen attached to it. 
• 3. antibodies surround it
• 4. macrophage ingests antibody-bacteria-antigen complex
• (ANTIBODY PROMOTES PHAGOCYTOSIS)

• 1. bacteria is in plasma of cell
• 2. antibody identifies bacteria/antigen
• 3. antibodies surround it with "complement"
• 4. lysis occurs
• 5. macrophage ingests destroyed parts
• (ANTIBODY ACTIVATES A COMPLEMENT CONNECTION, WHICH CAUSES OPSONIZATION AND LYSIS)

• 1. Nonspecific and occurs soon after infection.
• 2.Repeated exposure to pathogens does not improve innate immunity.

• 1.Specific and begins days or weeks after infection.
• 2.Repeated exposure to pathogens can improve adaptive immunity

• 1. our innate immune system searches for PAMPS (pathogen associated molecular patterns)
• 2. EXAMPLES:
• A. Lipopolysaccharide (LPS) is an example of a bacterial PAMP
• B. dsRNA is an example of a viral PAMP

• 1. a pathogen imposter: looks like pathogen but doesn't sicken host
• 2. pathogens covered with antigens that trigger immune response.
• Vaccine has similar antigen so body can have proper reaction to it before the real deal
• 3. APCs roam and find vaccine antigen. ingest and display the vaccine antigen. 
• (vaccine antigen is a PAMP)
• (APCs are dendritic cells, NK cells, and macrophages)

• 1. long extensions
• 2. are antigen presenting cells
• 3. will internalize and digest pathogens
• 4. display peptides from pathogens on surface to helper T cells
• A.. How is peptide brought to surface? By MHC (major histocompatibility complexes)
• a.MHC I: present antigens from inside the cells
• b.  interact with CD8 from cytotoxic T-cells
• MHC II: present antigens that are phagocytosed

--- interact with CD4 from helper T-cells

• 1. Searches for and destroys cells NOT displaying MHCs
• 2. 5-10% of the white blood cell population
• 3. Are activated by interferons (IFNs: are a group of signaling proteins made and released by host cells in response to the presence of several pathogens, such as viruses, bacteria, parasites, and also tumor cells) 
• 4. Respond to viral infections within minutes
• 5. release perforins and granzymes into host cells
• 6. can also Antibody-dependent cell-mediated cytotoxicity (ADCC): an effector cell of the immune system actively lyses a target cell, whose membrane-surface antigens have been bound by specific antibodies.

create pores in the membrane of target cells

activate apoptosis in target cells

• 1. Engulf and destroy foreign matter 
• 2. Will patrol for and pursue foreign matter (using chemotaxis)

• movement of a motile cell or organism, or part of one, in a direction corresponding to a gradient of increasing or decreasing concentration of a particular substance
• (like osmosis?)

• 1. release antimicrobial granules to aid in destroying pathogens
•    e.g., peroxidases, lysozymes, proteases

• 1. proteins that are produced in response to viral infection and initiate antiviral activity
• 2. > 300 IFN stimulated genes

• Example antiviral activity in response to dsRNA:
• PKR
• 2′5′-oligo(A) synthetase
• Rnase L

• 1. Programmed cell death
• 2. Can be activated by:
• a.Receptor binding
• b. Activation of PKR
• c. Changes in cell cycle progression
• d. Natural Killer cells

• 1. consists of more than 30 small proteins found in blood
• 2. one complement protein activates another in a cascade
• 3. opsonizes (dentifying the invading particle to the phagocyte) cells to enhance phagocytosis
• 4. Attracts macrophages and neutrophils 
• 5. destroys membranes
• a. MAC (membrane attack complex): Affects enveloped viruses

• 1. T and B cells
• 2. lymph supplies these to bloodstream
• 3. congregate in lymph nodes and through outgoing lymph vessels

• 1. CD4+
• A. helper T cells(T_h)
• B. stimulate T cell proliferation
• 2. CD8+ cytotoxic T cells (T_c)
• A. recognize MHC-1/antigen presentation
• B. secrete perforins and granzymes to destroy target cell

• APC's showcasing the antigen travel to where immune cells cluster , such as lymph nodes
• Effect:
• Naive T cells :specific to antigen recognize it as foreign and become activated
• Active T cells: (such as helper T cells) alert nearby T cells to the presence of the invader

• 1. epitope (binding area of antigen) is broken into accessible peptide 
• 2. epitope binds to MHC molecule from dendrite
• 3. T cell receptor (TCR) binds to epitope-MHC complex

• 1. When differentiated to plasma cells, can produce antibodies
• 2. neutralize or interfere with infectivity
• 3. Are antigen-presenting cells (APCs)
• 4. Activated by helper T-cells that recognize the antigen

IgM: induced shortly after infection

IgG: long-term protection; located in blood

IgA: located in secretions --- muscus, gastrointestinal tract, breast milk

• 1. When differentiated to plasma cells, can produce antibodies
• A. neutralize or interfere with infectivity
• 2. B cells work in concert with T_H2 cells

• 1. Recognize MHC-I/antigen presentation
• 2. Secrete perforins and granzymes to destroy target cell

cytokines

repeated exposure to pathogens results in maintenance of above

large (>2500 mw)

• intermediate = high
• high or low= low

particulate (not soluble) and denatured (not native)

slow release and bacteria

• enhanaces immune response
• helps it last longer, enhances macrophage

• ability to induce the immune reaction
• think about antibodies

don't want anything growing in vaccine

• alum hydroxide:
• 1. delays release of antigen
• 2. enhances macrophage uptake

prevent other microorganisms growing in solution, no bacteria growth

• keeps vaccine unchanged
• ex/ sterile water, saline, fluids containing protein

• 1. helps to grow vaccine antigens
• 2. found in influenza and yellow fever vaccines

• 1. used to inactivate bacterial products for toxoid vaccines
• a. toxoid vaccines: vaccines that use an inactive bacterial toxin to produce immunity
• 2. used to kill unwanted viruses and bacteria that might contaminate the vaccine during production
• 3. most formaldehyde is removed from vaccine before packaging

1. stabilizers in a few vaccines to help vaccine remain unchanged when exposed to elements such as HEAT, LIGHT, ACIDITY, HUMIDITY

• 1. mercury containing preservative that is added to vials of vaccine that contain more than one dose to prevent contamination and growth of potentially harmful bacteria
• 2. concern with link to autism
• 3. removed in some countries

• put in different host, virus mutates so much that when put back in different host, it reproduces so slowly that the different host is able to fight back.
• PRO's:
• a. cheaper
• b. cells do all the work, (more natural)
• c. higher antigen content following replication
• d. longer antigen persistence results in higher Ab responses
• CON's
• a. could possibly re-mutate back to the original

• 1. have been grown in culture and then killed using a method such as heat or formaldehyde.
• CON:
• 1. weaker response by the immune system than live viruses,
• 2. immunologic adjuvants and multiple "booster" injections may be required to provide an effective immune response against the inactivated pathogen.
• PRO's:
• 1. attenuated might pose risk for people with weakened immune systems so inactivated vaccine is better option

• 1. live over inactivated
• 2. protein vs. polysaccharide:
• 3. adjuvant:
• 4. genetic determinants: ability of Ag epitopes to be associate to large panel of MHC molecules -> (increases) liklihood of responses in the population
• a. gene polymorphisms most likely effect Ab responses

• 1. dendritic cells
• 2. NK cells
• 3. Macrophages

Nk cells, monocyes, macrophages, and neutrophils have Fc receptors for the Fc region of antibody so that the antibody attaches to it.

• 1. worms: can cause nk cells to activate naive cd4 t cells, which diff. into T H2 cells
• 2. viruses/bacteria cause dendritic cells to secrete IL-12 that activate nk cells to produce an interferon , naive t cell activated by both secretions , differentiate into Th1 cells

• 1. High mutation rate avoids specific antibody binding
• 2. Molecular mimicry: pathogen proteins are very similar to host proteins (our body cant detect them!)
• 3. Can synthesize a lot of pathogen proteins and exhaust immune system ....(too much to process)
• 4. Can dysregulate cell gene regulation, including those involved in the immune response
• 5. Can block cell signaling, such as apoptosis and complement system

• 1. caused by variola virus
• 2. best known for: being a large virus, inducing blistery rashes
• 3. poxviruses are species specific
• 4. symptoms: vomiting, diarhea

• 1. small host range
• 2. 1/2 bil. deaths in 20th century

• infects various mammals
• similar to cowpox and buffalopox
• mild rash and fever at day 0
• scab separates at 14-21 days

• small dosage of real thing to develop robust immune response
• ex/ chinese AD usage of blowing scabs up the nostrils

• 1. developed by Edward Jenner
• 2. more cautious (obviously)
• 3. used vaccinia virus for variola virus
• 4. Inoculated boy with cowpox, then smallpox. blister formed for cp but nothing occurred for sp!

• 1. developed by leslie collier
• 2. bifurcated needle
• 3. added peptone
• 4. created a heat stable vaccine

• 1. all fevers treated as smallpox
• 2. several guards posted at home
• 3. everyone within 10 miles vaccinated
• 1972: last administered to U.S. public

• live vaccine by dryvax strain
• over 200 mil doses stockpiled
• 2003 civilians and military personnel still vaccinated

• 1. high level immunity 3-5 years (decreasing immunity thereafter)
• 2. prevent or substantially lessen infection when given within a few days of exposure
• 3. >90% ppl vaccinated 25-75 years ago still maintain substantial humoral or cellular immunity against vaccinia
• a. antibody response remained relatively the same, T cell response declined w/ half life 8-15 years
• b. booster increased antibody, not CTL response

• 1. mild: arm receiving vaccination is sore, armpit glands sore, low fever, 1/3 ppl bad enough to miss work
• 2. Serious: 1/1000  ppl receive a vaccinia rash, or toxic or allergic rash
• 3. Serious: (14-52/10^6) serious skin rashes caused by widespread infection of skin, ppl w/skin cnoditions
• a. tissue destruction, leading to death
• b. inflammation of the brain

• 1. lab workers who handle smallpox
• 2. emergency response /healthcare workers
• 3. military personnel
• 4. ppl administering smallpox vaccine
• Post event:
• 5. ppl exposed to smallpox virus
• 6. ppl at risk of exposure for smallpox virus

• 1. ppl w/ allergy to vaccine components
• 2. <12 yrs, or over 65 yrs
• 3. skin conditions
• 4. pregnant women
• 5. breastfeeding women
• 6. weak immune system
• 7. steroid drops in eyes
• 8. ppl with poor health

can inoculate another body part such as your eye

• 1. ssRNA
• 2. bullet shaped virus (habdoviridae family)

define all

• 1. attaches to nicotonic acetylcholine receptors on muscle cells
• 2.
• 3. after replicating in muscle cells, virus is able to cross the synaptic cleft and bind to neural cell adhesion molecules (NCAM) on neuronal cells
• 3.b.
• 4. virus travels within axons in peripheral nerves up CNS, at rate of .5-15" per day
• 5.

• 1. raccoons
• 2. bats
• 3. skunks
• 4. cats

account for 17% of all rabid animals

• 1. 10 million postexposure vaccinations
• 2. 55,000 deaths world-wide
• 3. since 1990, over 90% of  human cases due to bats

• 1. occur w/in 20-60 days
• 2. "furious" -encephalitic (inflammation of the brain)
• 3. "dumb"-paralytic
• 4. Incubation: 18-21 days
• 5. before coma: 2-14days
• 6. death: 18+days after symptoms

• 1. hydrophobia
• 2. difficulty swallowing
• 3. agitation
• 4. hallucinations
• 5. hypersalivation
• 6. bizarre behavior
• 7. biting
• 8. violent contractions of diaphragm

• 1. lack of hydrophobia
• 2. lack of hyper -reactive behavior
• 3. lack of seizures
• 4. weakness and paralysis

• 1. headache
• 2. malaise
• 3. fever
• 4. anorexia
• 5. nausea
• 6. vomiting

• human diploid cell rabies vaccine
• Def: the virus is harvested from infected human diploid cells, MRC-5 strain, concentrated by 13 ultrafiltration and is inactivated by beta-propiolactone
• 3. from human cultured fibroblasts

• from fetal rhesus lung cell culture.
• 2. used in U.S. (only in michigan?)
• 3. RVA is adsorbed to AlPO4, it is liquid rather than lyophilized

• Purified chick embryo cell vaccine
• 2. sterile lyophilized vaccine obtained by growing the fixed rabies virus strain Flury LEP-25 in primary cultures of chick fibroblasts.
• 3. primarily used in US and germany

• 1. for high risk individuals
• 2. vaccine given intramuscularly in deltoid on days 0,7, 21, or 28

• 1. Do not cauterize the wound
• 2. Wash wound with 20% soap or 70% EtOH and virucidal iodine solution
• 3. On day 0, injected with HRIG and vaccine
• 4. Provides passive and active immunity
• 5.Additional vaccination days 2, 7, 14, 28

• developed the rabies vaccine from attenuating it from rabbits and using it on dogs and people hosts
• Roux one of his students

boy who was the first one cured from rabies by Pasteur

• 1. G protein(Glycoprotein or hairs on virus): neutralizing antibodies inhibit this protein from binding to receptors on cells
• 2. CD8+ CTL responses produced against N protein (ribonucleoprotein or strand that wraps around in virus and genomic RNA is encased)
• a. this destroys infected non-neuronal cells before virus entry into CNS

• igM: antibodies appear 4 days after first dose of vaccine
• igG: antibodies appear 7-14 dasy after first dose
• age is factor, >50 may respond less

takes several days for virus to replicate in muscle before reaching neurons

• 1. neutralization antibodies decrease after the first year
• 2. neutralizing antibodies are below acceptable levels after two years (results vary)
• 3. booster shot recommended after 1 year, see titer level

partial genetic vaccine against rabies for wild raccoons and coyotes

• small ssRNA virus
• icosahedral shaped
• nonenveloped
• acid-stable

• 1. portal of entry: mouth
• 2. person to person spread: oral-fecal route
• 3. infants most efficient transmitters of infection
• 4. incubation: 6-20 days
• 5. poliovirus : 3-6 weeks in stool
• 2 weeks in saliva

• 1. enters orally
• 2. replicates in oro-pharynx tonsils
• 3. also can replicate in intestinal area called peyers patches

• viral ligand = vp4
• attaches to receptor cd155

• Inactivated:
• 1. salk used formalin to kill virus
• 2. milzer used uv radiation to kill virus
• Attenuated
• 3. savin created 3 strains of live attenuated virus

1. injected CNS tissue of polio infected boy into two monkeys

• 1. Enders, Weller, Robbins: cultivation of polioviruses in non-nervous tissue
• 2. monkeys treated w/ picric acid , sodium alum nasal spray. abandoned on humans bc not effective
• 3. gamma globulin: dose of antibodies to prepare body for polio. deemed ineffective

• 1. grew polioviruses in monkeys
• 2. inactivated the virus with formalin
• tests on children showed favorable

• 1 licensed in 1955 by FDA
• 2. 70% effective in preventing poliovirus infection

• 1. developed opv (live attenuated poliovirus vaccines)
• 2. more appealing bc came closest to producing natural situation
• 3. longer lasting immunity
• 4. major concern: can they revert to wild type?

• 1. hard to gain suppor of another polio vaccine
• 2. trial in singapore
• 3. russian professor used seed strain of sabine and vaccinated 15 mil russian children. led to 100 mil russians with no effects

• 1. contains neomycin and streptomycin
• 2. shed in stool for up to 6 weeks following vaccination
• 3. immunity probably lifelong
• 4. NO LONGER used to avoid VAPP (vaccine associated paralytic polio)

• a. 4 doses of ipv at 2 months
• b. 4 doses of ipv at 4 months
• c. 1 dose of ipv at 6-18 months
• d. 1 booster of ipv at 4-6 years
• intramuscularly

• Cutter incident
• 1. manufacturer of salk vaccine did not adequately inactivate vaccine
• a. 260 cases of vaccine relateed poliomyelitis
• (family, vaccination, community)
• 2. did NOT change public confidence in vaccine
• 3. led to pharmaceutical GMPs

• 1. monkey rehsus cells
• 2. to chimpanzee cells
• 3. then to cultured cynomolgus monkey cells

• 1. administered , few drops orally
• 2. WHO supports in developing countries due to intestinal immunity advantages
• 3. live virus, can be transmitted oral-fecal route to unvaccinated ppl, creating secondary spread of vaccine

• 1. neutralizing IgG antibodies created for life
• 2. higher IgA antibody levels than for IPV
• CON: VAPP, 1/2.4 *10^6 cases, no longer recommended by 2000 in US

3. in 1960 rhesus monkeys ahd SV40, which can cause tumors in rodents. not sure if this causes cancers with ipv

• 1. two vaccines available
• 2. no animal reservoir
• 3. 3 attenuated serotypes are stable
• 4. OPV is inexpensive and easy to administer

• 1. poliovirus is a contagious and stable virus
• 2. use of IPV in tropical regions is problematic
• 3. IPV is inefficient in preventing the spread of virus
• 4. some resistance to mass vaccination
• 5. how can you tell if successful, only 1/100 ppl suffer from paralytic polio