Clinical Laboratory Methods PPT 12- Immunology & Serology updated and Revised 2024/2025, Exams of Clinical Medicine

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Clinical Laboratory Methods PPT 12-

Immunology & Serology u pdated and

Revised 2024/

What are the 2 immune responses? (__immunity) - ANSWER>>innate immunity acquired immunity

which immunity is non specific? which is specific and adaptive? (acquired or innate) - ANSWER>>1. non specific -> innate immunity

2. specific -> acquired immunity

What's an inflammatory response? - ANSWER>>response to infection or tissue injury

which immunity has barriers against general pathogens? - ANSWER>>innate immunity

What are the steps to an inflammatory response? - ANSWER>>Chemicals are released from injured or infected site Blood vessels dilate and allows neutrophils to pass into tissue Phagocytosis by neutrophils; causes pH change Monocytes migrate into tissues; now macrophages and cont phagocytosis

what are some signs of inflammation? - ANSWER>>redness, swelling, heat, pain, loss of function

NK cells are part of what cell class? - ANSWER>>class of lymphocytes

where are NK cells found? - ANSWER>>blood and peripheral lymph organs

Nk cells recognize and kill what cells? - ANSWER>>kill host cells that are coated with IgG

how do NK cells activate phagocytes? - ANSWER>>through the use of interferon

what are interferons? - ANSWER>>soluble proteins secreted by cells

Interferon is a cytokine and mainly the big reason it is used is to do what?** - ANSWER>>prevent viral replication

what else does interferon do? - ANSWER>>Elicit other cellular reactions Influence NK cells Also active in adaptive system as well as innate system

what is complement? - ANSWER>>collective group of proteins found in blood

True or False complement acts in both innate immunity and adaptive immunity systems\? - ANSWER>>TRUE

Activation of complement causes what to occur? - ANSWER>>activated cause complement cascade (3 pathways)

Classical is a type of complement pathway in which what happens? - ANSWER>>Classical: adaptive, activated C1 binds to antigen-antibody complexes that are formed in the body

Innate is a type of complement pathway in which what happens? - ANSWER>>Innate; microbial surfaces and plasma lectin binding (big idea: it binds to microbial surfaces)

Complement has many physiological effects. Name 2 - ANSWER>>Opsonization of microbes..makes them more easy to find and more readily recognized and more easily destroyed Promotion of phagocytosis Stimulate inflammation..stim. continued inflammatory response Stimulate cell lysis Membrane attack complex

what are the 2 components of acquired immunity? - ANSWER>>humoral (B cells) cell mediated (t cells)

if the pathogen gets passed innate immunity what does your body also have? (what other immunity system) - ANSWER>>adaptive immunity

adaptive immunity responds specifically to what? - ANSWER>>to antigens

what is the goal of adaptive immunity? - ANSWER>>to neutralize, detoxify, and eliminate

what is the main cell type of adaptive immunity? - ANSWER>>lymphocytes

in the adaptive immunity, macrophages are responsible for what? - ANSWER>>for process and presentation of antigens (antigen presenting cells)

adaptive immunity develops where? matures where? - ANSWER>>Develop in yolk sac, then fetal liver, then bone marrow (once patient is born) Mature in either bone marrow (B cells) or thymus (T cells)..

in humoral immunity, ___ are secreted by the B cells into ___ - ANSWER>>Antibodies (Ig molecules) are secreted by B cells Secreted into body's fluids ..(aka hemour)

how many stages of maturation are there in humoral immunity? what are they? - ANSWER>>3 stages of maturation Lymphoblast Prolymphocyte Mature lymphocyte

once the cells are mature, where do they go? - ANSWER>>Mature cells leave marrow for secondary lymph tissue: mainly spleen and lymph nodes

True or False Each B Cell develops specific receptor molecule for specific antigens - ANSWER>>true -Antigen reacts with committed B cell to produce antibodies

how are antibodies produced? (step by step of antibody production) - ANSWER>>B cell reacts with antigen Helper T cells produce cytokines Cytokines activate reactive B cell B cell divides and differenties Clones are produced and secrete antibodies..undergo clonal expansion where they replicate over and over

plasma cells are a type of __ cell - ANSWER>>effector

antibodies are immunoglobulins with what shape configuration? - ANSWER>>protein molecules in Y shaped configuration

what are the 2 different regions in the Y shaped configuration called? what does each region do? - ANSWER>>Antigen binds to variable regions Constant region is responsible for differing Ig classes

how many classes of Ig are there? what are they? - ANSWER>>5 classes of Ig IgM

IgG IgE IgA IgD

which Ig is the most abundant? - ANSWER>>IgG

which Ig is produced first? - ANSWER>>IgM is the first produced antibody

IgM: - ANSWER>>5 monomers 5% of circulating Ig Relatively large Responsible for activation of complement** First antibody type produced..to a antigen

IgG: - ANSWER>>Most abundant type of Ig Approx 75% of circulating Ig Remains in circulation longest Capable of entering tissues based on small size and produced during secondary immune response Responsible for neutralization of microbes and toxins..main job Opsonization of microbes..makes them more readily detectable and killed Complement activation Fetal and neonatal immunity by passive transfer..through colostrum etc

IgE: - ANSWER>>Very small amounts..in most patients Responsible in immediate hypersensitivity reactions Coating of parasites for destruction by eosinophils..comes in and binds parasites and flags them to be destroyed by eosinophils

IgA: - ANSWER>>20% of circulating antibodies..second most abundant! (first is 75% igG) Responsible for mucosal immunity

Protection of respiratory, intestinal, and urogenital tracts

IgD: - ANSWER>>Very low abundance Responsible for coating B cell surface antigens

what is neutralization? - ANSWER>>when antibody binds directly to antigen

the variable region binds to _____ the constant region binds to _____ - ANSWER>>1. binds to receptors on microbial surface

2. binds to macrophages and/or neutrophils

Antibodies react to prevent ___ or ___ of microbes into the body - ANSWER>>attachment or invasion

when antibodies bind to an antigen on a cell surface what happens? - ANSWER>>when they do this you get rupture/lysis of that cell

-can cause antigenic cell lysis

True or False when antigen binds to antibodies you can have reactions - ANSWER>>true ex. precipitate form, insoluble complex

true or false memory cells work in both parts of adaptive immunity - ANSWER>>true works in humoral and cell-mediated immunity

cell-mediated immunity is __ effector cells - ANSWER>>T effector cells

1. helper T
2. cytolytic or cytotoxic T

True or False

In cell mediated immunity, the cells develop receptors to specific antigens - ANSWER>>true

Helper T cells recognize _____ and release ___ - ANSWER>>recognize phagocytized antigen (APC) release cytokines (cause further phagocytosis and B cell differentiation)

Cytolytic T cells recognize ___ on surface and then cause ___ if infected cell - ANSWER>>antigen lysis

True or False The body can occasionally have self reacting lymphocytes - ANSWER>>True sometimes we can have lymphocytes that are programmed to react to self- antigen

*we can also have leukocytes that are supposed to undergo immunologic tolerance

what is immunologic tolerance? - ANSWER>>This is the body's way of saying, these are self antigens if you react tot hem you should die vs. them continuing to proliferate. this is the body's way to control self antigen

what is anergy? - ANSWER>>unable to respond

What is apoptosis? - ANSWER>>programmed cell death

*die when encounter self-antigens

what is self-ignorance? - ANSWER>>nonimmunogenic: ignored by naive lymphocytes

if have self antigen, the body is trying to tell them to not respond, not react, not stimulate it

self ignorance and immunologic tolerance are the body's ways to control ___ and ___ - ANSWER>>self antigens self reactions

what is negative selection? - ANSWER>>Negative selection destroys lymphocytes that have self receptors that cause self destruction. Occurs in bone marrow, thymus, and peripheral tissue

what are regulatory T cells? - ANSWER>>suppressor T cells, prevent self reactive lymphocytes from becoming effector cells

when self ignorance, immunologic tolerance, and negative selection all fail what happens? - ANSWER>>autoimmune dz occurs

what are the 4 types of immune disorders? (type 1, type 2, type 3, type 4) - ANSWER>>Type I: allergies, anaphylactic shock; mast cells release chemicals and cause immediate hypersensitivity..ex peanut allergies, bee stings etc

Type II: IMHA; diseases caused by destruction of own cells by antibodies..ex IMHA, type 1 diabetes

Type III: Immune Complex diseases; caused by precipitation of antibody-antigen complexes in vessels..ex. Lyme nephritis->inflammation in kidneys due to precipitated out lyme antibodies

Type IV: Contact hypersensitivity; t-lymphs against self-antigens in tissue..ex. Dermatological rxn

if you are not producing enough lymphocytes, phagocytes, monocytes and immunoglobulins what can you see as a result of this? - ANSWER>>can see secondary infections bc weak immune system

can see cancer-> escapes natural defense mechanisms

What is the difference between passive and acquired immunity? - ANSWER>>Passive: preformed antibodies passing to animal; usually short lived but immediate immunity..immediately active..ex colostrum from mom to babies, can be seen when give plasma to patients

Acquired: active resistance due to having disease and developing antibodies; usually due to vaccinations

what are the 2 forms of vaccinations? - ANSWER>>Attenuated: weakened live versions;

Inactivated: killed versions; generally safer.

(attenuated/inactivated) is longer lasting and more potent, mild risk

(attenuated/inactivated) no risk of causing disease, shorter duration, less potent - ANSWER>>attenuated

inactivated

What are VAFS? (whats it stand for) - ANSWER>>vaccine associated fibrosarcomas

VAFS are mainly due to what? - ANSWER>>often due to the adjuvants that are added to the killed versions

Adjuvants often added to killed vaccines to increase potency..killed are just proteins not as strong so come companies increase proteins/chem to increase

potency (increase cytokine and immune response by the body) and those chemicals may increase VAFS formation thought to cause sarcomas

True or False VAFS can occur in dogs and cats - ANSWER>>FALSE VAFS is in cats ONLY

where should we give vaccinations on cats? (where on leg) - ANSWER>>Give as low as possible on limb to facilitate removal of leg if sarcoma develops

EX. RR-rabies RF-distemper FIV/FELV-LF

why do we give vaccines as low as possible on the limbs? - ANSWER>>by giving vaccine lower on leg if sarcomas form they can remove leg easier

what is serology? - ANSWER>>detection and measurement of antibodies or antigen

how is humoral immunity testing done? - ANSWER>>Monoclonal antibodies produced by immunizing, collecting antibody secreting plasma cells, fusing with cells that propagate indefinitely Attach to only one specific part of an antigen..variable portion of antigen

*variable portion of antibody attaches to a specific part on the antigens

What does ELISA stand for? - ANSWER>>enzyme-linked immunosorbent assay

ELISA has (high/low) specificity for antigen ELISA has (high/low) specificity for antibody - ANSWER>>high-antigen

low-antibody

True or False If testing for antibodies there may be cross reactivity if they have been recently vaccinated even if they are not infected - ANSWER>>true result may be positive even if not infected

how do you get a positive results for an ELISA test? (antigen-antibody presence) - ANSWER>>Monoclonal Antibodies bind to antigen and this binds to enzyme labeled antibody. Chromogenic substrate is added to react with enzyme and produce color change if positive..you have a substrate that changes color ..if positive get a color change if antigen present get color change if no antigen no color change

*antibody binds with specific antigen. if binds then enzyme will bind with substrate and produce a color change

in competitive ELISA, what two antigens are competing? - ANSWER>>Patient antigen competes with enzyme labeled antigen

true or false in competitive ELISA, Intensity of the color change varies with concentration of patient antigen - ANSWER>>true If patient has higher amount of antigen then test substrate you are going to get a deeper color change

what is latex agglutination? (steps to procedure) - ANSWER>>Small particles of latex are coated with antigen Serum is added If antibodies present in serum they will cause agglutination...antibodies will bind to antigens and cause agglutination

___ is the type of serologic test that is done in old + pregnancy tests - ANSWER>>rapid immunomigration

what is rapid immunomigration? - ANSWER>>Colloidal gold is conjugated to specific antibodies If antigen is in patient sample it will bind to antibodies Migrates to outer area where second antibody is present Color change Also has control area where there should be a color change Two areas of color change is considered positive

Immunodiffusion is what is sued in ___ testing for horses - ANSWER>>coggins

what is immunodiffusion? - ANSWER>>Patient sample and antigen are placed in wells on agar plate As they diffuse into agar they form a line where the antigen-antibody complex precipitates

in immunodiffusion, if there is no line forming what does this mean? - ANSWER>>If no line forms there are no antibodies in the patient's serum

What is a radioimmunoassay? - ANSWER>>Similar to CELISA except uses radioisotope instead of enzyme Measure radioactivity

what is the difference between direct and indirect fluorescent antibody testing? - ANSWER>>Direct: measures dye coated antigen that combines with patient antibodies..tag it with fluorescent compounds an dsee if it will light up under a black light Indirect: measures when patient antibodies bind to test antigen and secondary anti-antibodies

One is looking to see if the patient has specific antibodies to a specific antigen

The other one is going to look at whether or not the patient has antibodies to an antigen

what is Coombs testing? - ANSWER>>Detects self-antibodies Incubate sample with antisera Reacts with IgG on coated RBCs Visible agglutination

Indirect detects circulating self-antibodies; serum of patient with RBCs from normal and causes agglutination

direct : will take RBC from my patient and mmix it with normal serum (bc normal serum should not have antibodies)

what is Coombs testing mainly looking for? - ANSWER>>Looking for agglutionation and to see whether or not patient has antibodies to self antigen

intradermal testing: - ANSWER>>Used for various allergens Extracts are injected intradermal Monitored for signs of hypersensitivity Raised welts, redness

antibody titers may distinguished between ___ and ___ - ANSWER>>active and prior exposure

antibody titer: - ANSWER>>take serum, dilute it to the point where there are no antibodies left in that

the more titer= the more dilute = greater dilution

the higher the titer = the (more/less) amount of antibodies = (active/prior exposure) - ANSWER>>more amount active infection

the lower the titer = the (more/less) amount of antibodies = (active/prior exposure) - ANSWER>>less amount prior exposure or vaccination

tuberculin test: - ANSWER>>(looking for) Specific cell-mediated reaction Animals infected with mycobacterium develop a specific delayed hypersensitivity response Injected intradermally If exposed there is a delayed inflammation Takes 24hr+ for t-cells to migrate to area

Molecular Diagnostics are used to analyze __ or ___ and can be used to do what?

• ANSWER>>Analyzing DNA or RNA

Can be used for: Classifying cancers Detecting genetic defects Verify pedigree..like those Dna kits Determine viral, fungal, bacterial infections

molecular diagnostics advantages vs disadvantages - ANSWER>>Advantages Increased sensitivity Increased specificity..^ bc bpth increased = generally really accurate Small specimen Not as affected by factors..dz state of patient generally doesn't affect this testing Faster turnaround

Disadvantages Technical expertise..cannot do in house High cost

What does PCR stand for? - ANSWER>>polymerase chain reaction

PCR is an amplification assay. what does this mean? - ANSWER>>makes copies of a small select region

how many steps are there to amplification? what are they? - ANSWER>>3 steps

1. Denaturation: DNA sample is heated to break apart strands, this serves as the template
2. Annealing: Temp is lowered to to bind primers to strands, signal beginning and end of section to be copied (replicated)
3. Extension: temp is raised and polymerase causes new DNA segment to be produced Process is repeated 25 to 30 times

what does RT-PCR stand for? what does it use? - ANSWER>>Reverse Transcriptase PCR Uses RNA instead of DNA

what is real time PCR? - ANSWER>>modified PCR

what are alloantibodies? - ANSWER>>self antibodies to RBC groups

True or False cats have circulating alloantibodies, dogs do not - ANSWER>>true

what does DEA stand for? - ANSWER>>Dog Erythrocyte Antigen

there have been 12 blood groups ID for dogs. which are the most clinically significant ones? - ANSWER>>Only DEA 1.1 and DEA 7 are considered to be clinically significant DEA 1.1 has the greatest response and causes the most serious effects

True or False dogs who are DEA 1.1 negative can get blood from 1.1 neg or 1.1 pos - ANSWER>>FALSE ideally 1.1 neg should only get 1.1 neg blood A negative dog should only get negative blood ideally -> bc if give positive blood body is going to be like I haven't seen these before and build antibodies to it and may have reaction to it if do second transfusion body already has antibodies to it and so bc repreated exposure body will kick those antigens out strat immune response

True or False dogs who are DEA 1.1 + can get 1.1 pos or 1.1 neg blood - ANSWER>>true can get both

the majority of cats are (A,B,AB) blood - ANSWER>>type A

True or False Cats so produce alloantibodies to different RBC groups - ANSWER>>true

Type A cats have (weak/strong) allo B antibodies - ANSWER>>weak

type B cats have (weak/strong) allo A antibodies - ANSWER>>strong

if you give type A cat type B blood what will occur? - ANSWER>>may have a mild rxn

if you give type B cat Type A blood what will occur? - ANSWER>>one drop can kill them bc type B has very strong allo A antibodies

Treu or False Type AB cats have A and B alloantibodies - ANSWER>>FALSE no alloantibodies

True or False cats do not need to best blood types before transfusion - ANSWER>>FALSE they should always be blood types prior to a transfusion!

Both congenital and acquired coagulopathies will benefit from administration of ____ or __ or ___ to replenish clotting factors. - ANSWER>>fresh blood, fresh plasma or fresh frozen plasma

Patients with severe thrombocytopenia may require transfusions of ___ or ___ or ___ in order to prevent life-threatening blood loss. - ANSWER>>fresh blood, platelet concentrate or platelet-rich plasma

what are some blood products? - ANSWER>>whole blood pRBCs (packed RBC) FFP (fresh frozen plasma) FP (frozen plasma) *natural colloids

whole blood:

pRBC:

FFP:

FP: - ANSWER>>Whole blood: contains rbc, plasma proteins (like albumin), and clotting factors. good for anything

pRBCS: good in cases of anemia..can be used interchangeably with whole blood FFP: all clotting factors and albumin. good for any coagulopathy and DIC..anticoagulation rodenticide, low albumin FP: good for pressure support and some coagulopathies..has albumin and only some not all clotting factors

...depednding on what you are treating you can use FP and FPP interchangeabely

is it possible to give a blood transfusion on a dog without first blood typing them?

• ANSWER>>Dogs do not have circulating alloantibodies so its possible to give dog a blood transfusion first without typing it and getting no response But second time must type them and see what they are before he subsequent transfusion bc if positive theier body is making antibodies to it so you don't want to give them the wrong blood again

what is the difference between major and minor crossmatch? - ANSWER>>MAJOR crossmatch = recipient serum mixed with donor RBCs to detect antibodies in the recipient against the donor's cells

MINOR crossmatch = recipient RBCs mixed with donor serum to detect antibodies in the donor's serum against the recipient's RBCs