ImmunoSero Flashcards
Network of organs
Immunology
Function of Immunology (RRRD)
Recognize
Respond
React
Destroy
PRIMARY Immune system organs
Bone marrow
Thymus
SECONDARY Immune system organs (5)
Spleen
Lymph node
MALT
GALT
Peyer’s patches
Site of B cell maturation
Produce hematopeitic stem cells
Bone marrow
Functionally similar to appendix in humans
Bursa of Fabricious
Cells that are not committed (lymphoid, myeloid, erythroid progenitors)
Stem cells
Site of T cell maturation and differentiation
Thymus
Site of T and B cell encounter
Secondary Immune System Organs
Houses the lymphoid (B and T cells)
MALT/ Mucosa Associated Lymphoid Tissue
Small pox vaccine; cross-immunity
Edward Jenner
DISCOVER Phagocytosis
Haeckel
EXPLAIN Phagocytosis - Cellular Theory of Immunity
Metchnikoff
Mendelian Inheritance
Gregor Mendel
Live, attenuated chicken cholera and anthrax vaccine
Louis Pasteur
Humoral theory of immunity
Emin Von Behring, Kitasata
Type IV Hypersensitivity Demonstration
Koch
Discover the complement
Jules Bordet
Antibody formation theory
Ehrlich
Immediate-Anaphylaxis Hypersensitivity
Portier, Richet
Discover the Opsonins
Almroth Wright
Large intestinal organ
Liver
Cells that can be found on: Chronic dse, tumor, organ donation
T cells
Compromised T cell immunity
HIV
D’george syndrome
Nezelof’s syndrome
Thymus is located in the:
Back of the sternum, near the heart
Largest external organ
Skin
Discover the C reactive protein
Tillet and Francis
Discover the Ag-ab binding
Marrack
Polio vaccine
Salk and Sabin
Oral polio vax
Sabin
Yellow fever vaccine
Reed
Clonal selection theory
Burnet
Discover the Monoclonal Abs
Kohler, Milstein
First immunoassay that used radioisotope
Radioimmunoassay
Discover the radioimmunoassay
Rosalyn Yallow
Year the thew T cell receptor gene was discovered
1985-1987
Antibody Diversity
Susumo Tonegawa
Discover the HPV vaccine
Frazer
T or F
HPV Vaccine is not allowed or given to men.
FALSE
Types of Immunity
Innate
Adaptive
Innate or Adaptive:
Non-specific
Natural
Without memory
Phagocytosis
Non- specific
Short-term
Innate
Innate or Adaptive:
Specific
Stimulated
With memory
T and B cell activation
Specific
Long-term
Adaptive
Pathogen Recognition for Innate Immunity
TLR or Toll-Like Receptors
Pathogen recognition for Adaptive Immunity
Memory cells (ex. Booster shot)
Mechanism of Immunologic Injury:
- Allergic or Anaphylactic
- innate
- Basophils and mast cells
Type 1
Mechanism of Immunologic Injury:
- Cytologic
Type 2
Mechanism of Immunologic Injury:
- Immune complex formation
- Ab attached to organs
Type 3
Mechanism of Immunologic Injury:
- Cell-mediated or delayed
- autoimmunity
Type 4
Responses of INFLAMMATION (3)
Vascular
Cellular
Repair/ Resolution
2 parts of Innate Immunity
External defense
Internal defense
The defense system that prevents entry of pathogens into the body
External defense
Defense system that kills pathogen that entered and established inside the body
Internal defense
pH of skin to prevent infection
4.8 to 6.4 pH
Normal flora in the skin
S.epidermidis
Normal flora in the oral cavity
Viridans strep.
Normal flora in the vagina
L.acidophilus
Lactate and fatty acid maintain skin at
5.6 pH
Soluble factors in the internal defense system
APR or Acute Phase Reactants
Structural barriers: Physical
Unbroken skin
Mucosal membrane
Tears
Sweat
Hair/Cilia
Structural barriers: Biological
Gut
Skin
Oral cavity
Vagina
Intestine
Gut produce: __________ that binds negatively charged surface if certain bacteria and penetrate to kill
Colicins
Bacteria that can be found in the intestine
Gram - anaerobic or bacteroides
Enterobacteriaceae
Cells for Cellular Factors (4)
Basophils and mast cells
Phagocytic cells
NK cells
Innate lymphoid cells
- Histamine
- Regulates T helper cell response to stimulate B cells to produce IgE
- Short life span
- Located in the BV
Basophil
- More histamine
- Has CP, ALP, Proteses
- For allergic rxn and ag presentation
- Larger
Mast cells
Growth factor for endothelial cells
VeGF or Vascular Endothelial Growth Hormone
- Vasoactive amine
- Dilate the BV locally
Histamine
Produced by in the BV for dilation and autocrine signaling
Nitric oxide
What happens to BV if dilated?
Will have SPACES
Heparin will control RBCs not to go out
Enzyme that destroys proteins
Proteases
How many IgE is needed to activate mast cells?
2 IgE
Leukotriene inhibitor
Montelukast
Arachidonic acid produces (2)
Prostaglandin
Leukotrienes
Ig for
- Th1
- Th2
- Th17 & 9
Th1= IgG & IgM
Th2 = IgE
Th17 & 9 = IgG ang IgM
Th1
Viral
Th17 & 19
Bacterial
Increasing amount of exposure
Desensitization
Given to px with anaphylactic shock
Epinephrine
- Principal phagocyte/ first to go to the site of infection
- Bacterial infx
Neutrophils
Other name for Neutrolphil
Segs
Segmenter
MIcrophage
2 population granules of Neutrophil
Primary/ Azurophilic
Secondary/ Specific
Primary granules contain:
MPO
Lysozyme
Elastase
Proteinase-3
Cathepsin G
Defensins
Destroys elastin
Elastase
T or F:
Proteinase-3 has anti-bacterial property
True
Secondary granules contain:
Lysozymes
Lactoferrins
Collagenase
Gelatinase
Respiratory burst components
T or F:
Respiratory burst components are stored in secondary but formed in primary granules.
True
Fat that is destroyed by macrophage
Mycolic acid
Leukocytes based on abundance
Neutrophil
Lymphocytes
Monocytes
Eosinophil
Basophil
Phagocytic cell:
- Regulation of immune response
- mast cell indirect phagocytosis
- increased in allergic rxn
- increased in parasitic infection
Eosinophil
Contents of Eosinophilic granules
Catalase
Lysozyme
Cytokines
Growth factors
Cationic proteins
Major basic proteins
Histaminase
Create a hole in helminths in basic pH
MBP
Largest cell in the peripheral blood
Monocytes or MAcrophages
Monocyte 1st type of granules components:
Peroxidase
ACP
arysulfatase
Monocyte 2nd type of granules components:
B-glucoronidase
Lysozyme
Lipase but no ALP
Interleukins released by the monoyctes during phagocytosis
1B
6
12
Interleukin that go to liver to produce CRP to ingest organism
Il-6
Interleukin that regulate temperature or stimulates fever
Il-1B
Interleukin that activate NK cells for anti-tumor activity
Il-12
Coat organism to be recognized
Opsonins
Two types of macrophages
M1 - Classically activated
M2- Alternatively activated
T or F:
Monocytes are lipid in nature.
True
What interferon activates M1
IFN-y
What Il activates M2 (2)
Il- 13 and Il- 4
M1 produces __, ___, ___ during phagocytosis
ROS
NO
Lysosomal enzymes
M1 produces ___, ____, ____, ____ during inflammation
Il-2
Il-12
Il-23
Chemokines
M2 produces growth factors and TGF-B for:
Tissue repair
Fibrosis
M2 produces Il-10 and TGF- B / Transforming growth factor - B
Anti-inflammatory effects
Most effective antigen-presenting cell and most potent phagocyte
Dendritic cells
Phagocytic cells
Neutrophil
Eosinophil
Monocyte or macrophage
Dendritic cells
Phagocytic cells for healing and repair
Macrophage and dendritic cells
MACROPHAGE: Lungs
Dust cells
Alveolar macrophage
MACROPHAGE: Liver
Kupffer cells
MACROPHAGE: Bone
Osteoclast
MACROPHAGE: Placenta
Hoffbauer cells
MACROPHAGE: Brain
Microgilial cells
MACROPHAGE: Kidney
Mesangial cells
MACROPHAGE: Connective tissue
Histiocytes
MACROPHAGE: Spleen
Littoral cells or splenic macrophage
MACROPHAGE: Blood
Monocytes
MACROPHAGE: Skin
Langerhans cells
MACROPHAGE: Synovium
Type A lining cells
cell eating cell
Phagocytosis
Steps in Phagocytosis (6)
Initiation
Chemotaxis
Recognition
Ingestion/ Engulfment
Digestion
Exocytosis/ Egress
2 types of Phagocytosis
Direct
Indirect
Direct phagocytosis via:
PPRR - organism
PAMP - WBC
Indirect phagocytosis via:
Opsonins
Most potent chemoattractant
C5a
Most potent opsonin
CRP
Physical contact bet. WBC and foreign cell
Initiation
Recruitment of immune cells to the site of infxn
Chemotaxis
Chemical messengers that aid the WBC movements
Chemoattractants or Chemotaxin
Chemoattractant
65a
C3a
CRP
Il-8
Squeezing out of WBC from blood vessel to tissue
Diapedesis
Process of Diapedesis
Rolling
Adhesion
Transmigration
Rolling:
L-selectin with bind to
Sialyl-Lewis X
Adhesion:
Integrin will bind to
E-selectin
T or F:
Lewis is produced in endothelial cells.
True
Direct phagocytosis:
- distinguish self from non-self
PPRR
Direct phagocytosis:
- present in the foreign organism
PAMP
PAMP: Gram +
Peptidoglycan
PAMP: Gram -
Lipoprotein
LPS
PAMP: Yeast
Zymosan
PAMP: FLagellae
Flagellin
Receptors in DIRECT phagocytosis
PPRR
PAMP
TLR/ Toll-like receptor
CLR/ C-type lectin receptor
RLR- Retinoic Acid - Inducible gene I-like receptor
NOD/ Nucleaotide-binding oligomerization domain receptor
Examples of opsonins
CRP
C3b
Antibodies
Light chain enhancer of activated B cell
NF-kB
Toll protein is discovered from
Fruit fly - Drosophila
CLR binds to ______ and ____ that are found in the fungal cell walls
Mannan
B-glucans
Forms Phagosome or vacuole
Ingestion or Engulfment
Lysosomal granules + Phagosome =
Phagolysosome
Defect in lysosomal enzyme
Chediak Higashi
Secondary Digestion Pathway
HMP or Hexose Monophosphate Shunt
Digestion of microbes by hydrolytic enzymes
Secondary digestion
Defect in NADPH Oxidase
Chronic Granulomatous Disease
2 secondary digestion process
Oxygen dependent
Oxygen independent
- Oxidative burst via HMP shunt
- Produce Oxygen Radicals ( Superoxide, Hydrogen Peroxide, Hypochlorite)
Oxygen dependent
- NADPH Oxidase
- Depolarization membrane
- Hydrogen and potassium will enter then alter pH then activate Proteases
Oxygen independent
Oxygen radicals that reacts with superoxide
Nitric oxide
Convert NADP+ to NAD+
NADPH oxidase
Convert Superoxide to Hydrogen Peroxide
Superoxide dismutase
Convert Hydrogen Peroxide to Hypochlorite
Myeloperoxidase
Damages cell membrane
Cathepsin G
Release debris outside the cell
Exocytosis or Egress
Toll-like receptors found on the cell surface
TLR 1 - Techoic acid
TLR 2 - Lipoproteins
TLR 4 - Lipopolysaccharides
TLR 5 - Flagellin
TLR 6 - Lipopeptides, Lipoteichoic acid, Zymosan
Toll-like receptors found on the endosomal compartments
TLR 3 - dsRNA
TLR 7 and 8 - ssRNA
TLR 9 - dsDNA
TLR 10 - unknown
- cytoplasm
- serves as intracellular sensors for microbial products
- inflammatory responses
Nod-like receptors or Nucleotide oligomerization domain
- cytoplasmic sensors
- production of type 1 interferon
Rig-1-like receptors or Retinoid-inducible gene
- Kiss of death
- First line of dense against cells that are: virally infected, infected with intracellular pathogen, tumor cells
NK cells
Large Granular Lymphocyte
NK cells or Natural Killer Cells
T or F:
NK cells are positive in CD16 (FcRy) and CD 56 (CAM)
True
T or F:
IFN-y and TNF-a produces cytokines when activated (NK cell)
True
Link between innate and adaptive immunity
NK cells
NK cells monitors potential target cells thorugh (2)
Inhibitory signal
Activating signal
Inhibitory signal:
Receptors which bind to MHC 1
CD49/ NKG2A
Activating signal:
Receptors (cancer cells)
CD16
NKG2D
Activating signal:
Receptors (viral infxn)
ADCC: CD16
ADCC
Antibody-dependent cell cytotoxicity
Site: DNA virus
Nucleus
Site: RNA Virus
Cytoplasm
Process: VIRUS
- “A-P-U-M-A-R”
Attachment
Penetration
Uncoating
mRNA
Assemble
Release
- Bone-marrow-derived cells
- with lymphocyte morphology but lack markers from lymphocytes
Innate lymphoid cells
Tissue signal: Type 1 effect
against intracellular oganism
Tissue signal: Type 2 effect
parasitic helminth and allergy
Tissue signal: Type 3
extracellular organism
Most potent antigen-presenting cell
Dendritic cells
Follows phagocytosis
Antigen Presentation
Present Ag to CD8+ cells
MHC Class 1
Present Ag to CD4+ cells
MHC CLass 2
Type of Signal (Antigen Presentation):
- CD 3 proteins
- Z chains
- TCR heterodimer
Type 1
Type of Signal (Antigen Presentation):
- CD28 (T cell) to CD 80/26 (Dendritic cells)
Type 2
Type of Signal (Antigen Presentation):
- IL-12
Type 3
Surveillance cells that checks the MHC Class 1
Nk cells/ T cell
Major Histocompatibility Complex Location
Chromosome 6p
MHC produces
Human- Leukocyte Anitgen (HLA)
T or F:
Major function of MHC is to aid in antigen presentation to T cells
True
MHC test of choice
Molecular methods (transplant institute)
Type of MHC:
- All nucleated cells
- Present to CD 8+
- Endogenous antigen
- Viral or cytosol antigen
- Ag: HLA A,B,C
- 3 alpha, 1 beta
MHC Class 1
Type of MHC:
- B cells, phagocytic cells (Macrophages, neutrophils, dendritic)
- Exogenous antigen
- Bacterial antigen
- Ag: DP, DQ, DR
- 2 alpha, 2 beta
MHC Class 2
Type of MHC:
- RBC
- Not capable of antigen presentation
- Ag: Benett Goodspeed
MHC Class 3
Disease = HLA present:
Goodpasture’s
Multiple sclerosis
DR2
Disease = HLA present:
SLE
Autoimmune thyroid disease
Dermatitis
Herpetiformis
DR3
Disease = HLA present:
Rheumatoid arthiritis
DR4
Disease = HLA present:
Ankylosing spondylitis or Ankyloarthritis or Bamboo Spine Disease
B27
No intervertebral disc
Ankylosing spondylitis
Disease = HLA present:
Celiac disease
Dermatitis Herpetiforms
B8
Disease = HLA present:
Psoriasis vulgaris
Cw6, B17, B13
Disease = HLA present:
Congenital Adrenal Hyperplasia
B47
Laboratory Test for MHC (Old methods)
Microlymphocytotoxicity Test
Mixed Lymphocyte Reaction
Newer detection methods for MHC
ELISA Crossmatch
Flow Cytometric Antibody Screen
Multiplex Immunoassay (LUMINEX)
- Tests for HLA Antigen
- Use purified HLA ag instead of lymphocytes
ELISA Crossmatch
- Use T or B lymphocytes or purified HLA ag
- Coat into microparticles
Flow Cytometric Antibody Screen
- Use Px’s HLA ag
- Mean Fluorescence Intensity
Multiple Immunoassay (LUMINEX)
LAD Score Method
Logarithm of the Odds score
Biologic Response Modifiers
Humoral Factors
4 main sources of BRMs from the mononuclear leukocytes
B lymphocyte
T lymphocyte
NK lymphocyte
Monocytes & Macrophages
T or F:
Monocytes & Macrophages secretes IFN-a, Il-1, TNF-a, GM-CSF, M-CSF
True
- Small soluble proteins
- Chemical signal functions to recruit other immune cells
Humoral Factors
- Polypeptide product of activated cells
- Control a variety of cellular responses
Cytokines
- First cytokine activity described
- Retention and accumulation of phagocytes at the sites of infxn
Migratory Inhibitory Factor (MIF)
MIF or Migratory Inhibitory Factor Examples
Interleukins
Tumor Necrosis Factor
Interferon
Transforming Growth Factor
Innate Immunity Cytokines
Chemokines
IFN Type 1 (a & B)
Il-1,6,10,12,15,18
TNF
Adaptive Immunity Cytokines
IFN- y
Il-2,4,5,13
Lymphotoxin
TGF-B
Types of Cytokine:
- affects same cell that secreted it
Autocrine
Types of cytokine:
- secreted by nearby cells
Paracrine
Types of cytokine:
- secreted by the circulatory system
- affect distant celss
Endocrine
Massive overproduction of cytokine leading to shock, multi-organ failure, death
Cytokine Storm
Rapid screening of bacterial infx secondary to covid
Procalcitonin
- COVID Receptor
- Induce to produce more cytokine
ACE-2 Receptor
Functions of Interferons (3)
Enhance specific gene expression
Inhibit cell proliferation
Augment Immune effector cells
Type of interferon:
- Leukocyte IFN
- activates NK cells
- inhibits viral replication
Type 1; IFN-a
Type of interferon:
- Immune IFN
- Secreted by T cells, Th1, NK cells
Type 2; IFN-y
Type of Interferon:
- Fibroepthelial IFN
- secreted by dsRNA-induced fibroblast
Type 1; IFN-B
Principal mediator of Acute Inflammatory response to Gram -
Tumor Necrosis Factor
effective against Gram + except streptococcus
Beta- lysin
Site of production of TNF
LPS-activated macrophage
Recruit and activate phagocytes to kill microbes
TNF
TNF Concentration:
- acute inflammation
Low concentration
TNF Concentration:
- systemic effect of inflammation
Moderate concentration
TNF Concentration:
- causes clinical and pathological abnormalities
Large concnetration
TNF-a
Cachetin
TNF-B
Lymphotoxin
- Released by Platelets during coagulation
- heat stable
Beta-lysin
Multicolony stimulating factor
Il-3
- Peptides and proteins
-Serves as signal molecules - Functions through separate receptor
- Leukocyte is the site of production
Interleukin
- glycoproteins
- increase rapidly during infxn, injury, tse trauma
- promote phagocytosis and limit destruction through proteolytic enzymes
APR or Acute Phase Reactants
T or F:
APR is initiated and sustained by cytokine Il-1, Il-6, TNF-a
True
Half-life of APR
2-4 days but CRP has 5-7 hrs
APR Protein:
- for opsonization and complement activation
- main substrate is Phosphocholine
CRP Or C reactive protein
APR Protein:
- Activate monocytes and macrophages
- Increase in chronic inflammation, atherosclerosis, cancer
Serum Amyloid A
APR Protein:
- general plasma inhibitor of proteases
- inhibites eleastase
a-1-antitrypsin
APR Protein:
- general plasma inhibitor of proteases
- inhibits elastase
- regulates elastase, Il-1B, Il-6, TNF-a
a-1-antitrypsin
AAT deficiency
Premature emphysema
Liver disease
APR Protein:
- Nine serum proteins
- Present and mediates inflammation
Complement
APR Protein:
- Antioxidant
- Binds irreversibly to free hemoglobin
Haptoglobin
APR Protein:
- cleaved by thrombin to produce fibrin clot
- promotes aggregation of RBC and platelet by making blood viscous
- Normal value = 200-400 mg/dl
Fibrinogen
APR Protein:
- Principal copper-transporting protein
- Binds 6 cupric ions per molecule
- Wilson’s disease
- Normal value: 20-40 md/dl
Ceruloplasmin
Overall body reaction to injury or invasion by an infectious agent
Inflammation
T or F:
The main purpose of inflammation is to attract cells to the site of infection for phagocytosis
True
Roles of inflammation (IAS)
Initiate
Amplify
Sustain
Primary process in inflammation
Localized inflammation
Final process of inflammation
Resolution and repair by Fibroblast
- Early stage of infection
- Shirt term
Acute Inflammation
- Prolonged infection
- Long term
Chronic inflammation
Resolution and repair makes fibroblast proliferate resulting to
Total repair
Abscess formation
Granuloma formation
Cardinal signs
Rubor - erythema
Calor - hear
Tumor - edema
Dolor - pain
Functio laesa - loss of function
Pressure sensor
Pacinian corpuscle
Protein complex that recognizes products of dead cells
Ex. Uric acid - by product of purine
Inflammasome
Components of Inflammasome
Sensor protein
Adapter
Enzyme caspase-1
Activate caspase-1
Il-B
Third line of defense
Adaptive immunity
Types of adaptive immunity
Humoral - abs
Cell-mediated - T cells and B cells
Cells of adaptive immunity
T cells and b cells
T or f:
Th1 = intraellular microbes
Th2 = helminthic parasites
Th3 = extracellular bacteria, fungi
True
- 60 o 80% lymphocyte fraction
- for cell-mediated immmunity
- selected and matures from cortex to medulla
T-cells
Develomental stages of Tcells
Double negative
Double positive
Mature T cells
Activated T cells
- No CD4 and CD8 markers
- Proliferate due to Il-7
- Gene arrangement for TCR
Double Negative
Selection of allele in 1 chromosome
Allelic Exclusion
Express both CD4 and CD8
Double Positive
Types of Double Positive
Positive selection - retain thymocytes with functinal tcr
MHC Selection - selects thymocytes
Negative selecttion - apoptosis of cells
Colonal deletion- eliminates clone
- Either CD4 and CD8 is present
- Thelper and T cytotoxic cells
Mature T cells
T cells with 2/3 population
T-helper cells
T cells with 1/3 population
T cytotoxic cells
- Produce CD25 (Il-2 receptor) to secrete cytokines enhnce B cell ab production
- Prevent autoimmunity
Activated T cells
Sub-population of Tcells
Th9 = Il-9 produtiion (extracellular bacteria)
Th17 = Il-17 and Il-22 (inflammation and bone destruction)
- inverted CD markers
- 1:2 ratio
AIDS
- For humoral mediated immunity
- in the bone marrow (stromal cells forming niches)
B cells
Phases of B cell development
Phase 1 - antigen independent
Phase 2 - antigen dependent
Phase 3 - differentiation of plasma cells
Phase 1 cells
Pro-B cells -heavy and light chain
Pre-B cells - heavy chain and u chain
Immature B cells - complete IgM
Mature B cells- surface IgD
Elimination of B cells bearing self-reactive receptors
Central tolerance
Halmark of adaptive immune system
Pre-existing diversity of receptor for Ag
Not an end stage cells
Surveillance cell
Lymphocyte
Remain in the spleen to quickly respond to blood-borne pathogens
Marginal zone b cells
Migrate to secondary lymphoid organs
Follicular cells
Most fully differentiated lymphocyte
Plasma cells
Activators of lymphocytes
Monoclonal - antigens
Oligoclonal - superantigens
Polyclonal - mitogens (mitosis =allow proliferation of lymphocytes)
B cell mitogen
Lipopolysaccharide
T cell mitogen
Concanavaline A
Phytohemagglutinin
Both T and B mitogen
Pokeweed mitogen
Laboratory Identification of Lymphocytes (3)
Density Gradient Centrifugation - Ficoll- Hypaque
Roswell Park Memorial Institute Medium - RPMI 1640
Flow cytometry - gold standard
- E-rosette Test
- CD2 receptor on sheep RBC
- End product: cytokines
- Location: Paracortical region
T cell
- Surface Ig Detection
- IgD and igM on the surface of naive B cells
- End product: Abs
- Location: Cortical region
B cell
Forms of Acquired Immunity
Active Natural - infxn
Active Artificial - vaccination
Passive Natural - in-vivo transfer
Passice Artificial - infusion of plasma/serum
- Proteins
-Binds with Ag - Site of Production = Plasma cells, spleen, lymph nodes
- Determinant = PARATOPE
Antibody
Antibody Structure
2 light chains - kappa or lambda
2 heavy chains - gamma, alpha, delta, epsilon, mu
Change in class of antibody
Isotype switching
Structural regions of Antibody (5)
Variable = amino terminal end
Constant = carboxy terminal end
Hinge = bet. Ch2 and ch3; PROLINE
Fc = fragment of crystallization; WBC binding site
Ch2 and Ch3 = complement binding site
Enzyme digestion (2)
Papain = 1st disulfide bond, 3 fragments
Pepsin = 2nd disulfide bond, 2 fragments
Unit of sedimentation rate of Immunoglobulin
Svedberg
Heavy chain differences
Isotypes
Minor variation
Allotype
Variable part of LC and HC
Idiotypes
pH for Ab gamma region
8.6
Production of monoclonal antibody by a hybridoma cell (immortally producing abs)
Hybridoma technology
Plasma cell + Myeloma cell
Hybridoma cell
Surfactant used for Hybridoma Technology
PEG or Polyethylene Glycol
Medium used for Hybridoma Technology
HAT or Hypoxanthine - Aminopterin - Thymidine
Formerly used potentiator for Hybridoma Technology
Sendai virus
Antibody Isotypes (5)
IgG
IgA
IgM
IgD
IgE
- Anamnestic response antibody
- Activate complement classical pathway
- Most abundant in SERUM
- most efficient in PRECIPITATION
- Can cross placenta
- Opsonin, viral neutralization, ADCC
IgG
Types of IgG
IgG1 = cross placenta
IgG2 = cannot cross immediately
IgG3 = fix complement
IgG4 = cannot fix complement; destroyed in the spleen
- Secretory components
- B cell production (MALT)
IgA
Types of IgA (2)
Monomeric IgA = blood
Dimeric IgA = secretions
- Primary reponse antibody
- Activates complement classical pathway
- Acute infection indication
- With joing chain or J chain
- Macroglobulin
IgM
Types of IgM
Monomeric IgM = surface of naive B cell
Pentameric IgM = plasma
- Surface of naive B cell
- Extended hinge region
- For B cell activation, maturation, differentiation
IgD
- Most heat labile
- Reagenic antibody/ Homocytotropic antibody
- Binds with mast cells, eosinphils, basophils
IgE
Antigen- Antibody Interaction:
- Association constant
- Univalent antigen
Affinity
Antigen- Antibody Interaction:
- Overall binding
- Multivalent antigen
Avidity
Antigen- Antibody Interaction:
- Test negative
- Abscence of homologous ag
Specificity
Antigen- Antibody Interaction:
- Test positive
- Presence of homologous antigen
Sensitivity
Antigen- Antibody Interaction:
- Bind similiar but not identical isotope
Cross Reactivity
Antigen- Antibody Interaction:
- PRIMARY Interactions (3)
Vanderwaals = hydrophobic
Ionic and hydrogen bonds = hydrophilic
Covalent bond
Antigen- Antibody Interaction:
- SECONDARY Interactions (2)
Precipitation = antibody + soluble antigen
Agglutination = antibody + particulate antigen
Stages of Agglutination (3)
Sensitization
Lattice Formation
Effect on Tissue
Factors that affect Ag- Ab binding
Ag - Ab ratio = Pro zone, zone of equi, post zone
pH = 6.5 to 7.5
Length of incubation
No. of antigens
Location of antigens
Centrifugation
Potentiators = LISS, PEG, Bovine albumin
Rouleaux and True agglutination = Add NSS for pseudo agglutination
Antibody isotope
Temperature = IgM (RT) , IgG (high temp)
Antibody generators
Antigen
- Antigenic determinant = Epitope
- 2 types= complete and incomplete (need haptens)
Antigen
Antigens capable of inducing immiune response
Immunogen
Forms of Antigens (3)
Autologous = induce autoantibody formation
Homologous = antibody production specific to antigen
Heterologous = cross-reaction, reacts with antibody it did not induced
- Small molecules
- Immunogenic when paired with high MW carrier
Haptens
- Added to vaccine
- Enhance immune response
- Ex. Squaline, MF-59, Freund’s complete adjuvant, Alum Precipitate
Adjuvants
- Antigens that cancactivate T cells by nonspecifically binding to MHC molecule
- Do not require anitgen processing
- Cytokine storm
Superantigens
Detectable range of test system
Linearity
Dengue serotype that can cause hemorrhagic fever
Dengue serotype 1
Traits of Immunogen (5)
Macromolecule size = > or = 10 to 100 kDA
Foreigness = Non-self (Lens protein, sperm, brain, thyroglobulin)
Chemical composition = Proteins and polysaccharides
Ability to be processed and presented with MHC
Route and dosage = IV and Intraperitoneal
- Least immunogenic organ
- No blood supply
Cornea
- Most immunogenic organ
Bone Marrow
Properties of Immunogens
T cell-independent = stimulate B cells directly, Ag processing is not required
T cell-dependent = Need T cell recognition, Ag processing is required
Induces cytolytic destruction by forming MAC to make hole in the cell membrane
Complement System
COMPLEMENT SYSTEM:
Inactivated at ____ for ____ mins
Must be used within _____ hrs
56 degrees celsius
30 mins
4 hrs
COMPLEMENT SYSTEM:
If not used within 4 hours, heat at _____ celsius for ______ mins
56 degrees
10 mins
Complement Pathways (3)
Classical = ab is attached to ag
Alternative = activating surfaces like LP
Lectin Pathway = lectin initiates binding of MBL/ Mannose Binding Lectin
Used for screening for bacterial infection
Procalcitonin
Regulatory Proteins (5)
C1 inhibitor / C1INH = detaches C1r and C1s from C1
Factor I = cleaves C3b and C4b
Factor H = cofactor to factor I to inactivate C3b
C4 Binding Protein = cofactor to factor I to inactivate C4b
S protein/ Vitronectin = prevents attachment of C5b67 to cell membrane
Disorders resulting from complement deficiency
C1 inhbitor = Hereditary Angioneurotic Edema
C1, C2, C4, C7 = SLE- like syndrome
C5 - C8 = Neisseria infection
C3 = Severe recurrent infection
C2 = most common deficiency
C9 = no known disease
- Exaggerated response
- harmless antigens that results to tuissue injury
- Influenced by environment and gene
- Treament: Anti-histamine, docingestant and corticosteriod
Hypersensitivity reaction
Monoclonal anti-IgE blocks the binding site of IgE to mast cells and basophils
Omalizumab
- Preferred method of screening type 1 in hypersensitivity reaction
- Smaal mamt of allergen is injected in the skin
- Wheal or flare reaction within 20 mins
In Vivo Prick Test
- Alternative test instead of In Vivo Prick Test
- Unable tol tolerate skin testing
Noncompetitive Solid Phase Immunoassay
Types of Hypersensitivity Reactions
Type 1 = allergic or anaphylactic
Type 2 = cytotoxic
Type 3 = immune complex deposition
Type 4 = delayed
Immune mediators for Types of Hypersensitivity Reactions
Type 1 = IgE
Type 2 = IgG or IgM
Type 3 = IgG or IgM
Type 4 = T cells
Immune Mechanism of Types of Hypersensitivity Reactions
Type 1 = IgE sensitized mast cells and basophils
Type 2 = ADCC destruction
Type 3 = Ag - Ab complex
Type 4 = Ag - sensitized Th1 cells to relase cytokines
Clinical examples for Types of Hypersensitivity Reactions
Type 1 = Anaphylaxis, allergic rhinitis, asthmas, food allergies, urticaria
Type 2 = HTR, AIHA, HDF, Drug reaction, Myasthenia gravis, Graves, Good pasteurs
Type 3 = Serum sickness, Arthus reaction, SLE, RA, Drug reaction
Type 4 = contact dermatitis, tuberculin and anergy skin test, hypersensitivity pneumonitis
Tests for Types of Hypersensitivity Reactions
Type 1 = In vivo skin test
Type 2 = DAT (HTR, AIHA, HDFN)
Type 3 = - Ag from animal source
Type 4 = Skin test, PPD testing (48-72 hrs)
- Failed tolerance results to
Autoimmunity
Autoimmune Diseases = Autoantibody
SLE = Anti-DNA, Anti- nuclear, Anti- ribosome, Anti-DNP
Primary biliary cirrhosis = Anti-mitochondrial
Chronic Active Hepatitis = Anti- smooth muscle antibody
Hypothyroidism = Anti- TPO, Anti-microsomzl Anti-thyroglobulin
Hyperthyroidism = Anti- TSH receptor, Anti-thyroglobulin
Goodpasteur’s Sydrome = Antiglomerular Basement Membrane (Edema)
Wegener’s Disease = c-ANCA
Churg - Strauss Sydrome = p-ANCA
DM Type 1 = Anti-insulin, Anti- Beta cells
Addison’s Disease = Abs against adrenal glands
Multiple sclerosis = Anti-myelin sheath ab
Myasthenia gravis = Anti-acetylcholine recptor
Pernicious anemia = Anti-parietal cell
Rheumatoid arthritis =Rheumatoid factor
ITP = Anti-platelet
Pemphigus vulgaris = Anti-desmosome
Bullous pemhigoid = Anti- hemidesmosome
Laboratory diagnosis for Autoimmunity
LE preparation
Rheumatoid factor
Immunochromatography
Immunofluorescent staining
Circular dsDNA substrate in anti-dsDNA detection
Crithidia lucilliae
Immmunofluorescent staining results:
Fluorescence patterns
Diffuse = anti-dsDNA and anti- Histone
Nucleolar = anti- RNP
Speckled = most common; anti-RNP and anti-smith
Peripheral = anti-DNA and anti-lamins
Centromere = anti- centromere; CREST (Calcinosis, Reynaud’s, Esophageal dysmotility, Sclerodactyly, Telangiectasia
Transplant immunology (3)
Allorecognition
Graft vs. Host Disease
Graft rejection
Allrecognition types
Autograft = same individual
Xenograft = 2 individuals, diff species
Allograft = 2 individuals, same species
Sygeneic graft = identical twins
Immunocompetent lymphocytes induced to immunocompetent individuals
GVHD
Type of graft rejection
Hyperacute = within minutes, humoral
Accelerated = 2 to 5 days, cell mediated
Acute = 7 to 21 days, cell mediated
Chronic= > 3 months, cell mediated
Immunopathologic damage to the new organ = > 3 months, immune complex disorder
Uncontrolled growth of normal cells
Neoplasia
Types of neoplasia
Benign = non-cancerous
Malignant = cancerous (-oma)
Neoplasia:
- Epithelial cell
Carcinoma
Neoplasia:
- connective tissue
Sarcoma
Neoplasia:
- bone marrow
Leukemia or Lymphoma
Protein seen in multiple myeloma
Bence Jones Protein
Spread of cancer cells
Metastasis
Tumor markers:
ONCOFETAL ANITGENS (2)
CEA = gastrointestinal, colorectal
AFP = hepatoma
Tumor markers:
CARBOHYDRATE ANTIGENS (4)
CA125 = Ovarian
CA15-3 = Breast
CA19-9 = Pancreatic, gastric
CA72-4 = Gastric
Tumor markers:
ENZYMATIC MARKERS (2)
PSA = Prostate
ALP = Bone
Tumor markers:
HORMONES (3)
Beta-HCG = Testicular
Calcitonin = Medullary thyroid
Gastrin = Gastric
Tumor markers:
ONCOGENES (3)
BRCA-1 and 2 mutations = breast
Her2neu = breast
CYFRA21-1 = lung
Tumor markers:
OTHERS (4)
Bombesin = oat cell
IGF-1 = pituitary
Il-2 = leukemia
NMP/ Nuclear Matrix Protein= bladder
Depleting immune system
Immunodeficiency
Immunodeficiency classification (2)
Primary = inherited (genetics)
Secondary = other factors (aging, stress, chronic disorders, viruses, lifestyle)
Immunodeficiency:
Phagocytic cell Deficiency (5)
CGD = defect incytochrome B
MPO def = defect in HMP shunt, decreased conversion to hypochlorite
G6PD def = genetic impairment of Neutophil’s aerobic system
Chediak higashi = genetic abnormal fusion of neutrophil granules (Albinism, photosensitivity, giant platelets)
Lazy leukocyte syndrome = (Job syndrome, Tuffsin def, Actin dysfunction)
Immunodeficiency:
B cell immunodeficiency (4)
X-linked Bruton’s Agammaglobulinemia
Common variable hypogammaglobulinemia
Selective IgA deficiency = most common congenital immunodeficiency
Neonatal agammaglobulinemia
Immunodeficiency:
T cell immunodeficiency (2)
DiGeorge syndrome = abnormal development of thymus gland
Nezelofs syndrome = athymic px
Immunodeficiency:
Combined T and B cell deficiency (3)
SCD = decreased T and B cells
Wiskot- Aldrich = Triad (Thrombocytopenia, immunodeficiency, eczema)
Bare lymphocyte = defect in MHC Antigen expression (Class 1 and 2)
Study of fluid component of blood (Ab)
Serology
Most frequent encountered specimen for serology
Serum
Commonly used Pipette in serological testing
Micropipettes
Tubes used for serological testing
Red top sterile tube / Gold top tube
Make less concentrated solution from a reagent
Dilution
Types of dilution
Simple
Compound
Serial = use titer (ab strength); common: doubling dilution
Proportion of people who have the disease
Sensitivity
Proportion of people who do not have the disease
Specificity
Probability that positive screening test has the disease
Positive predictive value
Probability that negative screening test does have the disease
Negative predictive value
- Ab excess
- Remedy: dilution
Pro zone
- Ag excess
Post zone
- Ag closely resemble one another
- Ab formed against one will react with the other
Cross reactivity
Removal of bound abs
Elution
Attachment of unbound abs
Adsorption
Sensitive than slide testing
Tube testing
Rapid reading and only need small volume of specimen
Slide testing
Agglutination grading:
- no agglutinates
0
Agglutination grading:
- tiny agglutinates, turbid
w+
Agglutination grading:
- small agglutinates, turbid
1+
Agglutination grading:
- medium-sized, clear
2+
Agglutination grading:
- Large agglutinates, clear
3+
Agglutination grading:
- One solid aggregates, clear
4+
Causes of false reactions in agglutination:
False positive reaction (7)
Overcentrifugation
Contaminated glasswares
Autoagglutination
Saline stored in glass bottles
Cross-reactivity
RF, Heterophile ab
Delay in reading
Causes of false reactions in agglutination:
False negative (7)
Undercentrifugation
Inadequate washing of cells
Delay in testing
Incorrect incubation temp
Insufficient incubation time
Prozone phenomenon
Failure to add AHG rgt
Agglutination rxns (7)
Direct
Indirect
Reverse passive
Agglutination inhibition
AHG mediated
Viral hemagglutination
Hemaglutination inhibition
- Natural carrier of Ag
- Febrile agglutination test
- ABO Forward typing
Direct agglutination
Febrile agglutination test (2)
Widal = salmonella ab
Weil felix = ricketssia ab
- Artificial carrier antigen
- Carriers = Polystyrene latex, bentonite, beads, charcoal
- ASO latex agglutination test
Indirect agglutination
- Artificial carrier of ab
- CRP Latex agglutination test
Reverse passive agglutination
2 stages of agglutination inhibition test
1st = add soluble rgt. ab
2nd = add ag-coated latex particle
AHG-mediated agglutination types (2)
Direct = in vivo, spx= rbc
Indirect = in vitro , spx= serum
Precipitation by Light measurement (2)
Nephelometry = light scattered
Turbidimetry = light blocked
Passive immunodiffusion types (2)
Single = only ag diffuses
Double = both ag and ab diffuses
Single immunodiffusion types (2)
Linear = James Oudin test
Radial = Fahey- Mckelvy (Kinetic) & Mancini (End point)
- Double immunodiffusion
- Double angular diffusion
- Ag (outer) , Ab (inner well)
Ouchterlony Double Diffusion
Result patterns for Ouchterlony Double Diffusion Test (3)
Identity = common epitope
Non-identity = no common epitope
Partial identity = single spur
General steps for Electrophoresis (3)
Separation = electrophoresis
Staining = Amido black, ponceau S, coomassie brilliant blue
Densitometry = band quantification
Separation Electrophoresis (2)
Zone Electrophoresis = 5 bands
High resolution electrophoresis = 12 bands
- One stage electrophoresis
- Single ID + Electric current
- Rocket precipiti
Rocket electrophoresis
- One stage electrophoresis
- Countercurrent
- Double ID + Electric current
- Precipitin line
- Ag moves to anode (+)
- Ab moves to cathode (-)
Counterimmunoelectrophoresis
Two stage electrophoresis (3)
Classic Immunoelectrophoresis
Immunofixation electrophoresis
Crossed 2D immunoelectrophoresis
- Electrophoresis + Immunodiffusion
- Trough
- Precipitin arc
Classic immunoelectrophoresis
- Electrophoresis + Immunodiffusion
- Gel
- Precipitin bands
Immunofixation electrophoresis
- Electrophoresis + electrophoresis
- 2nd gel
- Precipitin rockets
Crossed 2D immunoelectrophoresis
Marker of ag-ab binding
Label
Examples of label in Labeled Immunoassay
Enzyme
Fluorescein dye
Radioisotopes
Steps in Labeled Immunoassay
Separation = separate bound and free reactants
Detection = label dependent
Physical methods for Labeled Immunoassay: SEPARATION
Decantation, centrifugation, filtration
Solid phase vehicle for labeled immunoassay (5)
Polystyrene tubes
Microtiter tubes
Glass or polystyrene beads
Magnetic beads
Cellulose membranes
Label for Labeled Immunoassay : DETECTION (3)
Enzyme = Spectrophotometer
Fluorescein dye = Fluorometer, flow cytometer, spectrophotometer
Radioisotopes = Gamma scintillation counter
- Cheap and readily available
- Ag or ab detection use
- Commonly used labels = Alkaline phosphatase (Bovine intestine), Horseradish peroxidase (Horseradish), B- galactosidase (E.coli)
Enzyme aim,unoassay or ELISA
Noncompetitive EIA
Indirect = detects abs , solid phase= ag, direct concentration proportion , label AHG
Direct = detects ag, solid phase= ab, direct concentration proportion
, label Ab against ag
Sandwich = detects ag, solid phase= ab, direct concentration proportion , label AHG
- Detects ag
- Solid phase = ab
- Label = ag
- INVERSE concentration proportion
Competitive EIA
- Detects ag
- Solid phase = Unattached abs
- Labels = Abs
- Direct concentration proportion
Immunozymetric
- Typically used label = FITC, Tetramethyrhodamine, Phycoerythrin, Europium, Lucifer yellow
- Direct = rgt is labeled ab and speciment is ag
- Indirect = rgt is ag and labeled ag and specimen is ab
Fluorescence Immunoassay
Most popular radio immunoassay label
I 125
- First type immunoassay developed
- Radiation hazard
- Forms: Non-competitive, comptetitive, immunoradiometric
Radioimmunoassay
Quantitate total IgE
Radioimmunosorbent assay (RIST)
Antigen specific IgE
Radioallergosorbent Test (RAST)
- Emission light caused by chemical reaction
- Labels = Luminol, Acridium esters, Ruthenium derivatives, Nitrophenyl oxalate
- For cardiac marker, hormones, Vit D level, total IgE
Chemiluminiscent Immunoassay
Unexpected fall of analyte concentrstion due to ag excess
Hook effect
- uses electrochemical compound
- Solid phase = magnetic beads
- Light measured by photomultiplier tube
Electro-chemilumiscence Immunoassay
Two types of Nucleic Acids
DNA = deoxyribose sugar, double stranded
RNA = ribose sugar, single stranded
Central Dogma of Life
Replication
Transcription
Translation
Hybridization techniques
Southern blot
Microarray technology
Fluorescent in situ hybridization
Amplification techniques
Target amplification = PCR, Transcription mediated amplification, Ligase chain reaction
Probe amplification = standard displacement amplification, cleavase/ invader technology
Signal amplification = branched DNA, Hybrid capture assays
Whole genome amplification
- Amplifies tiny quantities of nucleic acid up to detectable levels
- Reagents = combination freshly prepared reagents
PCR or Polymerase Chain Reaction
Reagents of PCR:
- medium to suspend reagents
Water
Reagent of PCR:
- maintains pH
Buffer
Reagent of PCR:
- cofactor to enzyme
MgCl2
Reagent of PCR:
- Building blocks of DNA
Nucleotide
Reagent of PCR:
- starting point
Fore primer and reverse primer
Reagent of PCR:
- stabilize temperature (from Thermus aquaticus)
Taq polymerase
Steps in PCR: (3)
Denaturation = heat to 95 degrees to separate dsDNA to single strands
Annealing = cool to 52 degrees from primers to bind to complimentary sequence on each DNA strands
Elongation = 72 degrees, heat-stable DNA polymerase binds to primer syntehsize new strand of DNA
- Measures multiple properties of cells suspended in a moving fluid medium
- Identify and enumeratevvarious cell population
Flow Cytometry Analysis
Laboratory tests for Syphilis Serology (2)
Direct test
Non- treponelmal test
Species of Treponema (4)
T. pallidum subsp pertenue = YAWS
T. pallidum subsp endemicum = BEJEL
T. carateum = PINTA
T. cuniculi = RABBIT
Physical process of contact and adhesions wherein aggregates form karger-size clusters called flocs or flakes
Flocculation
Clinical stages of Syphilis (4)
Primary = hard chancre, blood
Secondary = conylomata lata, blood
Latent = no s/s, blood and csf
Tertiary = Gummata’s lesion,csf
Detects anti-cardiolipin
Non-treponemmal test
- Quali or quanti slide flocculation test
- Spx: Serum and CSF
- Read microscopically
VDRL
VDRL requires serum inactivation (2)
Heat serum for 30 mins at 56 degreees
Use serum within 4 hours
Reagents for VDRL (3)
0.03% cardiolipin
0.9% cholesterol
0.21% lecithin
Gauge of needles for Hamilton syringe (VDRL)
Qualitative = 18g (60 + or - then 2 drops)
Quantitative = 19g (75 + or - then 2 drops) or 23g (100 + or - then 2 drops)
Antigen that stabilizes Ag and inactivates complement (RPR)
EDTA
Na2HPO4
Thimerosal
Charcoal
Choline chloride
RPR:
Place serum-ag mixture in a rotator for _____ min @ 100 rpm
8 min
100 rpm
RPR:
- Uses plastic-coated disposable card with ______ circles instead of glass slide
10-18 mm circles
RPR:
- Gauge no. of needles
- Drops of Ag suspension/ml
20 gauge
60 + or - 2 drops
Old to new world
Smallpox
New to old world
Syphilis
Treponemal tests for Syphilis (2)
T. plallidum Immobilization Test
FTA-Abs
- Trepnemal test for syphilis
- Specimen = Serum
- Reagent = Motile bacterial suspension
- Interpretation = POS >50% immobilized, NEG <50, DOUBTFUL 20-50%
T.pallidum Immobilization Test
- Treponemal test for syphilis
- Specimen = Serum
- Reagent = dead T.pallidum in slide (Nichol’s virulent strain)
- Label = FITC- AHG
- Sorbent = Reiter strain
- View under UV light
FTA-Abs
Agglutination Tests (4)
TP Hemagglutination
Microhemagglutination
Hemagglutination Treponemal Test for Syphilis
TP- Particle Agglutination
Agglutination Test:
- Reagent = Tanned SHeep RBC coated with Ag from Nichols strain
- Confirmatory test = Indirect hemagglutination
TP Hemagglutination or TPHA
Agglutination Test:
- Reagent = Formalinized tanned sheep rbc coated with ag from Nichol’s strain
- Performed in microtechnique
Microhemagglutination or (MHA) TP
Agglutination Test:
- Reagent = Glutaraldehyed- stabilized turkey RBCs
- Automated version of TPHA
Hemagglutination Treponemal Test for Syphilis
Agglutination test:
- Reagent = Gelatin particles (instead of RBC) sensitized with T.pallidum
- Indirect Agglutination Test
TP- Particle Agglutination
- Retrovirus containing RNA and Reverse Transcriptase
- Target cell = CD4+ T cells
HIV Serology
Type of HIV :
- Formerly Human T-cell lymphotropic virus type III, Lyphadenopathy- associated virus, AIDS- associated retrovirus
- Causative agent of AIDS in US and Europe
HIV - 1
Type of HIV:
- Endemic in West Africa
- Less pathogenic
- Lower rate of transmission
HIV - 2
3 major routes of HIV transmission
Intimate Sexual Contact
Parenteral from body fluid/blood
Perinatal from infected mother (Vertical transmission)
Responsible for binding to CD 4 receptor of T cells (HIV)
gp120
First antibody to appear (HIV)
Anti-p24
HIV:
- Screening test
ELISA
HIV:
- Confirmatory test
Western blot
- Positive = + at least 2 bands
- Bands = p24, gp41, gp120/160
Western blot (HIV)
HIV:
- Ratio of CD4:CD8
1:2 or 0.5:1
- CD4 count in AIDS
<200 ul
Confirmatory test for HIV in Philippines
RHiVDA or Rapid HIV Diagnostic Algorithm
HIV:
- Test used 5 days from initial contact of infective agent
NAT
HIV strains (2)
T-tropic X4) strain - HIV infecting T cells (lymphocyte strain)
M-tropic (R5) strain - HIV infecting both T cells and macrophage (monocyte strain)
HIV Transmission %
- Blood transfusion
- Sexual intercourse; receptive anal intercourse
90%
6-8%
Syphilis Serology
RPR - screening test
FTA - confirmatory
ELISA - either screening or confirmatory
HIV Test:
- Gold standard = Immunophenotyping w/ Flowcy
- HIV AIDS CD4 count = <200 cell/mm3
CD4 T cell Enumeration Test
Screening tests forHIV Antibody Detection (3) “E A D”
ELISA
Agglutination test
Dot- blot testing
Confirmatory test for HIV Antibody detection (2) “W I”
Western blot
Immunofluorescence
Window oeriod before antibody is detectable
P24 Antigen detection
Determine viral load and development of drug resistant strain
HIV NAT
Quantitative test for HIV Nucleic Acid
Viral Load Test
HIV test preferred method for infants, hildren younger than 18 months
PCR
HIV Generation
First = detects ab to HIV 1 only
Second = detects ab to HIV 1 and HIV 2
Third = detects HIV ab of different Ig class including IgM
Fourth = detects HIV 1 and HIV 2 ab and p24 antigen
HIV Structural Gene (3)
Gag or Group Antigen Gene
Pol or Polymerase
Env or envelope
HIV Structural Gene = Product
- p25, core proteins ( p6, p9, p17, p24)
Gag or Group Antigen Gene
Products of Pol/ Polymerase gene (4)
Reverse transcriptase (p66, p51) = transcribes RNA to DNA
Integrase (p31) = inserts viral DNA to host
Protease (p10) = cleaves protein precursors
RNase = slipts DNA to RNA again
Products of Env/ Envelope gene (3)
gp160 = cleaved to form gp120 and gp41
gp120 = binds to CD4 on T cells
gp41 = transmembrane protein
Markers of Hepatitis B infection (6)
HBsAg = active infx
HBeAg = high degree of infectivity
IgM Anti-HBc = current or recent infxn, core windor period
IgG Anti-HBc = lifelong marker
Anti-HBe = recovery markerv
Anti-HBs = immunity to Hepatitis; Protective marker: > or = to 10 miu/ml of serum
Markers of Hepatits A infxn (4)
IgM anti-HAV = incubation period and early phase
IgG anti-HAV = Immunity to Hepa A
Total anti-HAV = Immunity to Hepa A
HAV RNA = HAV in clinical, food, water samples
Other names of Hepatitis:
- Hepa A
- may be transmitted by clotting factors
Infectious hepatitis
Other names of Hepatitis:
- Hepa B
Serum Hepatitis
Complete HBV that causes infxn
Dane particle
Other names of Hepatitis:
- Hepa c
Non-A, Non-B hepatitis
Post transfusion hepatitis
- Increased ALT
- Positive Anti-HBc
Surrogate Test for HCV
Positive Anti-HCV
Specific test for HCV
Tests for Streptococcal Serology (3)
ASO
Anti-DNase B Test
Streptozyme Test
_____ fold rise in titer to be considered significant (Streptococcal serology)
4 fold
Streptococcal Serology:
- 5 in 1 test = slide agglutinating screening test
- Reagent = Sheep RBCs coated with streptococcal extracellular products
Streptozyme
Streptococcal serology:
- Latex agglutination test = indirect or passive agglutination
- Tube test = neutralization or inhibition
ASO or Anti- streptolysin O titer
ASO tube testing units used:
Todd units = streptolysin rgt istandard is used
UI = WHO international standard
Normal= <200 todd unit
- Detects Ab capable of preventing DNase from depolymerizing DNA
- Principle = Neutralization
- Reagent = DNA-metho green substrate
- Grading= 4+ unchange color (positive), 0 toal loss of color (negative)
Anti- DNase B
Kissing disease caused by EBV
Infectious Mononucleosis
Atypical lymphocytes in EBV infxn
Downey cells
Infectious Mononucleosis Serology:
- Cross react with a group of similar antigen
- IgM usually appears during acute phase of ifxn
Heterophile Antibody
Heterophile Ab of IM:
- React with
- Absorbed by
- Not absorbed by
- React with = Horse, Ox, sheep rbcs
- Absorbed by = Beef rbcs
- Not absorbed by = Guinea pig kidney cells
IM Serological Tests (3)
Paul-Bunnel Screening Test
Davidsohn Differential Test
Monospot test
- IM serologic test
- Hemagglutination, detect heterophile ab
- Reacts with sheep rbc
- Normal titer: < or = to 56
Paul Bennel Screening Test
- IM serologic test
- Differentiates HA associated with IM, Serum sickness or Forssmann Ag
- Agglutination = Sheep rbcs
- Absorbed by = Guinea pic and Beef rbc
- Perform if Paul Bennel Screening test result is > 56
Davidsohn Differential Test
- IM serologic test
- Rapid differential slide test
- Requires absorption of px serum
- Horse rbc is agglutinated by heterophile ab of IM
Monospot test
Detects ab to Salmonella, Brucella, Tularemia
Widal test
Salmonella Serology:
- Clinical significant titer
> or = to 160
Antigens in Salmonella Serology (3)
O antigen = LPS
H antigen = Protein
K or Vi antigen = Polysaccharide/ capsular or virulent
Salmonella serology:
- Carrier specimen
Fluid from gallbladder (1 yr.)
Causes spotted fever and typhus
Rickettsia
- Test for detection of ab in rickettsia
- Four fold rise in titer or 1:160 is significant
Weil Felix Test
Antigens in Rickettsia serology (3)
OX-K
OX-2
OX-19
Ricketssia Serology: ORGANISM
- Epidemic typhus
- OX-19 (+)
R. prowazekii
Ricketssia Serology: ORGANISM
- Rocky mountain spotted fever
- OX-19 or OX-2 (+)
R. rickettsia
Ricketssia Serology: ORGANISM
- Scrub typhus
- OX-K (+)
R. tsutsugamushi
- Stool antigen test
- With endocscopy or No endoscopy
H. pylori serology
H. pylori serology:
- With endoscopy (3)
H. pylori culture
Histological exam
Urease biopsy test
H.pylori serology:
- No endoscopy (4)
Urea breath test
Enzyme immunoassay for bacterial ag in STOOL
Molecular test
Serum EIA
- Associated with cold agglutinins
- Auto p anti-I (blood banking)
M. pneumonia serologic test
Serologic test for ______:
- Thick smear = quantitation
- Thin smear = identification
Malaria
Immunochromatography for Malaria (2)
OptiMAL Assay
MalaQuick Stanby Marlarial test
Immunochromatography for Malaria:
- detects parasitic LDH (100-200 parasites/ul blood)
- detects variable parasite
- distinguish bet. spp through detection of different isoforms of PLDH
OptiMal Assay
Immunochromatography for Malaria:
- Detects P. falciparum Histidine Rich Protein (HRP)- 2 Antigen
MalaQuick Standby Test for Malaria
T or F:
H. capsulatum is a fungi.
True
Serological test for Histoplasma:
- 2 precipitin band of diagnostic significance
H line = most specific, found up to 2 yrs after recovery “HUMANA”
M line = found in px with active infx or past infxn or px who had recent skin test “MASASAKTAN KA PA”
Serological tests for Influenza (2)
Hemagglutinin = required for entry of virus (H1 to H18)
Neuramidase = for release of virus from cells (N1 to N11)
Mutations of Influenza (2)
Antigenic shift = occurs every 10 yrs, sudden in onset
Drift = occur at the site if the virus as it replicates, pre-existing abs
Most common cause of congenital infection
CMV or Cytomegalovirus
- Uses Frei test for Lymphogranuloma Vinerium
Chlamydia trachomatis
- Agent = Toxoplasma gondii
- Utilize Sabin-feldman dye test
Toxoplasmosis
- Utilizes Casoni skin test
Echinococcosis
