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Culture Documents
Microbiology
IMMUNOLOGY
Get reaction second time stung by bee; pre-existing IgE antibody specific for bee
venom is made on first sting.
Rash
Hives
IgE binds (Fe portion) to basophils and mast cells (Fe receptors).
Paratope - Antibody Combining site; the part of the antibody that binds to the
antigenic determinant.
Cell membrane –
No sterols except mycobacterium
ssDNA
ssRNA
Sigma factor - Specificity for promotor site recognition resides with this factor of
bacterial RNA polymerase
Mutation in the deletion of 1 base pair in the middle of a structural gene coding for a
polypeptide –
Results in changes in several amino acid residue within that polypeptide.
Ouchterlony test –
IgA
O
IgG O O O IgM
Anti-
IgG
Infection of a sensitive bacterial culture with a temperate bacteriophage might result in:
Lysis of some of the cells
Lysogenic (phage) conversion of some of the cells
Transformation –
In bacteria uptake naked DNA from solution into the cell.
Bacteria could acquire transposons
(No F factor needed)
Transduction –
Requires a bacteriophage, (NOT a sex or fertility plasmid)
Requires bacterial virus
Co – transduction –
Map bacterial genes by random breakage of chromosome during phage
lytic growth.
Conjugation –
Can result in passage of phage DNA from donor to recipient [F (fertility)
factor].
Transducing DNA –
Found only in one cell within a colony arising following an abortive
transduction event
Lysogenization –
Phage to prophage requiring ATP and ribosomes.
Example is diphtheria toxin.
λdgal –
Virus formed by aberrant excision of a prophage.
Hfr formation –
Integrates F into the chromosome.
During bacterial conjugation, acts as a donor of chromosomal DNA.
Cannot transfer prokaryolic genes among bacteria by meiosis nor mitosis.
Phage chromosomes –
Some contain only single-stranded RNA, some may be so small they have
genetic information sufficient for only 3 or 4 protein molecules.
Some plasmids –
May not be able to initiate bacterial conjugation
May be carried within the head of a bacteriophage
Generalized transducing particles – Contain bacterial genes (but not phage genes)
Antibody and macrophages (not cytotoxic T-lymphocytes, nor Natural Killer cells) –
Most important in host defense against extracellular invaders
Mal Ser E. coli grows on : Glucose and 1 amino acid Serine media.
The mouse –
Class 1 MHC: H-2K and H-2D
A nude mouse has no thymus
The mouse MHC is called H-2
Syngeneic mice are genetically identical
HLA-D –
Is called the immune response region in man.
It codes for antigens Important in cell-cell communication (Not code for
class I Ag’s)
It’s antigens are present on β-cells.
(does NOT code for certain complement proteins)
May prevent Graft vs. Host disease which can result from bone marrow transplantation –
Match MHC antigens of host and donor before transplantation
Pretreat bone marrow to remove mature T-lympocytes prior to
transplantation.
(Not remove all lymphoid cells and precursors from the marrow prior to
transplantation,
not use bone marrow from the mother for the transplantation)
MHC antigens are codominantly expressed – every cell that expresses antigen expresses
both alleles.
T-cells see antigen in the context of MHC antigens.
SRBC receptor:
CD2, T11, T cells. (NOT Fe receptor).
Class - Isotype
Suppressor T-cells –
Regulate immune responses.
Express CD8+ (human) or Lyt 23+ (mouse) antigens.
Exert effects on helper T-cells, and B-cells to inhibit immune response.
CD3+, CD2+
(NOT activated by helper or inducer T-cells.)
Helper T-cells –
CD4+
CD3+
CD2+
Lyt 1+23(-).
B-Lymphocytes –
Triggered by:
Antigen binding to surface immunoglobulin,
Activated T helper cells.
Helper factors IL-1, IL-2, and BCGE.
Macrophages –
Have a role in induction and amplification of immune response by:
Presenting antigen in an appropriate form to B and T-cells.
Expressing receptors for C3b, Fe part of IgG,m and lymphokines on their
membranes.
(NOT secreting lymphokines, and NOT by expression of antigen-specific
receptors on membranes.)
Class I MHC –
Cytotoxic T-lymphocyte (CTL).
Both B-cells and T-cells.
Class I antigens called Serologically Defined (SD).
Present on mast cells.
H-2K, and HLA-A.
Associated with beta2-microglobulin on surface.
T-Suppressor cells recognize antigens with MIIC class I molecules.
(Remember 1 x 8 = 8) CD8+
Class II MHC –
(Remember 11 x 4 = 8) CD4+
Delayed type hypersensitivity T-cells (DTII)
Helper T-cell
Helper inducer
T-suppressor inducer cells
B-cells, macrophages and activated T-cells
Gene products are HLA-D
Lymphocyte Defined (LD)
Class II MHC genes:
Are expressed on B-cells
Are involved in cell-cell communication
Are called lymphocyte defined or LD genes
Are important in antigen presentation
C5a and LTB4 - Directly chemotactic for neutrophils (NOT LPS, NOT 1L-2).
Neutrophils accumulate at inflammatory sites.
Overall –
Macrophages release IL-1 excites T-cells
release IL-2 which increases Cytotoxic T-lymphocytes, Natural
killer cells, BCGF (B-cells growth factor), and T-cells.
MHC complex –
Involved in graft rejection
Cell communication
Host defense.
Anaphylatoxins – C#a, C4a, and C5a (not C9a, Ba, C6a, C2a nor C1q)
Complement activators –
Ag-IgG1 complexes
Ag-IgM complexes
Bacterial endotoxin
Fungal cell walls
(not neutrophil surfaces)
BCGF,
Antigen, IL-2
Antigen-specific helper factors
IL-1: Is produced by macrophages
Is a genetically unrestricted and an immunologically nonspecific factor,
Is a polypeptide cytokine and is stable at temperatures (-750C to +560C)
and pH (3 to 11).
Known as LAF (lymphocyte activating factor),
Excites T-cells for IL-2 production, reaction to injury and other purposes.
Induces differentiation of T-lymphocytes by increasing metabolism.
(Not only effects T-lymphocytes)
(NOT MHC restricted
NOT heat labile at high temp.)
“Normal” regulation of an immune response, the following events may occur to dampen
the response:
Induction of anti-idiotype antibodies
Production of high affinity IgG antibody to remove antigen from the
system.
Helper cells – required for the activation of the following effector cells:
DTH and CTL (not macrophages and NK)
Properdin –
In alternate complement pathway, stabilized C3Bb.
CD4+:
Helper inducer,
Suppressor inducer,
Helper cells,
TDTH cells
In normal person, the number of CD4 lymphocytes is twice the number of
CD8 lymphocytes (ratio of 2:1)
(NOT suppressor effector, not TCTL)
(Not suppressor)
TI antigens - (T-Independent)
Are often polyclonal B cell activators.
Excite B-cell to produce mainly IgM antibodies, and have
little or no anamnestic (secondary) response to second stimulation.
TD antigens – (T-Dependent)
Can be IgE and IgA antibody-eliciting antigens.
Excite B-cells to make IgM antibody.
Generates anamnestic response to second stimulation (because antibody
switches to making IgG)
Antibody response, both T and B-cells have antigen specificity.
(Cell-cell contract is NOT essential).
The effects of IL-2 on a precursor CTL that has not yet seen antigen in the context of
MHC:
NONE, since IL-2 receptors do not appear until after specific stimulation.
“Altered-self” means –
The self-cell is altered (not the MHC antigens)
Myeloperoxidase deficient patients can still produce H2O2 • O2, and •O1I.
Marcophages –
Present antigen to the helper T-cell.
Phagocytose and kill some pathogens.
Remove immune complexes from the circulation.
Release II,-12
Important host defense mechanism because:
Process antigen and present antigen with MHC Class II (1a) molecule.
Produce cytokines, and an immune response.
Have nonspecific effector functions for specific DTH (NOT CTL).
Have important role as APC (antigen presenting cell).
Survive after phagocytosis (Neutrophils do NOT).
Activated Macrophages –
Have enhanced phagocytic activity.
Increased intracellular microbicidal activity.
Increased release of toxic produts (e.g., TNF).
Increased size and lysosomal content.
Macrophages –
Are involved in:
Presenting antigen to B and T cells
Induration seen in DTH skin test,
(NOT involved in immediate wheel/flare skin test type I hypersensitivity)
Are the major cell type used as host cells by: amastigotes of cutaneous
leishmaniasis.
Adaptive immunity –
Atopic allergy
Anamnestic response
Immunoglobulins
Specific antibodies,
DTH
(NOT interferons, NOT complement)
Innate Resistance –
Comp;e,emt
Monocytes
Macrophages
Natural killer cells
C-reactive protein
(NOT natural antibodies, Not DTH)
Mature B-cell –
When it can respond to immunogen by making antibody
IgD and other immunoglobulins (IgM), expressed on its surface
(Not when gene rearrangement of chain or express dId)
B-lymphocytes-
Development begins in fetal liver,
Surface IgM, IgG, IgA
(NOT increased with MHC molecules nor IFN)
B cell stimulated with DNP-BSA can cooperate with T-cell stimulated with ovalbumin
to produce an anti-DNP antibody response to DNP ovalbumin
In a 51Cr release assay, tumor cells release their 51Cr because they have been killed by a
cytolytic effector cell..
Thymus –
Cortex is mostly immature lymphocytes, therefore cells with TdT and
CD1+
Cyclosporin A –
Biological response modifier useful in heart, kidney, and bone marrow
Transplantation.
Inhibits T helper cell function.
Prevents IL-2 synthesis
TdT is present on –
Pre-B and pre- T cells
(not on immature B cells, nor medullary thymocytes)
Tumor cell or virus infected target cell lysis by specific cytotoxic T lymphocytes
requires: Cell-cell contact and specific recognition by T-cell receptor / T3
complex of CTL.
No antibody.
Divalent cations Mg 2+ and Ca2+ (Not complement)
Some antibodies against tumor target cells may block CTL killing of target cells, but may
help K cells to kill target.
NK cells –
Are distinct from classical T and B cells.
Increased by IFN and IL-2
Are primarily LGL.
Are induced by tumor cells.
Have a degree of specificity more than macrophages but less than T. cells.
(do Not require natural antibodies to kill target cells.)
(NOT induced by natural antigens.)
(NOT restricted by class I or II molecules)
(Are NOT thymus derived cells.)
IFN - (Interferon)
Increase NK activity, therefore used in immunotherapies,
Have anti-proliferation activity against tumor cells,
Immunoregulatory activities.
(NOT lymphokines)
Codominant expression –
Cyclosporin A –
Inhibits helper T-cells.
Prevents IL-2 synthesis
(Not a potent suppressor T-cell activator, not inhibit B-cells)
Western Blot –
Used to confirm HIV antibodies.
Known HIV antigens separated using electrophoresis.
Goal is to confirm a positive ELISA.
Positive bands when antibodies from patient serum binds to HIV antigens.
T cell Receptors –
Both alpha and beta chains, have variable and constant domains.
Recognize antigen in the context of MHC class I or classes II.
(MHC Class II or la antigen do NOT fix complement)
(CTL doesn’t recognize and kill specific virus-infection target cells
because CTL recognize mainly virus antigens in the context of MHC class
II antigen.)
B cell tolerance-
May occur if a single signal is received by a B cell.
is easier to induce in immature B-cells than mature B cells.
May be mediated by anti-idlotypic Ab.
May be either low zone or high zone tolerance.
Immunodeficiency -
Result of Malignancy and aging.
(NOT from vaccination and Increased intellectual activity.)
Immunodominant –
The most immunogenic determinant of a complex antigen.
Idiotype-anti-idiotype network.
Affects immunogenicity of antigen by complexing with antigen.
(Does NOT provide specific antibody to arm suppressor T cel Is.)
Ig classes can be distinguished based on their HEAVY chains (NOT light chains).
Opsonization
(Not Incr. vasodilation, not binding to basophils)
When IgG is digested with pepsin the following fragments are generated:
F(ab)2 which have two antigen binding si sites fragment. (Fragments will f ix
complement.)
IgG molecule:
The light and heavy chains are identical.
The carbohydrate is in the constant region of heavy chains.
(The hinge region does not join heavy and light chains.)
The IgG and the fragments of IgG were isolated from anti.- sheep red
blood cell antiserum: Agglutination occurs if IgG and sheep red blood cells
F(ab)2 and sheep red blood cells
(not Fab and sheep rbc, not Fc and sheep rbc)
Serum IgM:
Has J chain
Is a pentamer
(Does NOT have 2 Ab combining sites per molecule,
Not fix complement less efficiently than igG)
Highest titer in a hemagglutination assay in Inst agglutinated tube: (spread out not dot)
Serum IgM and membrane IgM have different properties for the following-
Size of heavy chain,
Molecular weight,
Number of I chains.
(Not size of light chains)
Idiotypic determinants.
Can induce anti.-idiotype Ab’s.
Are found in variable regions.
(Not found In constant .“ )
(can NOT produce anti.-allotype Ab’s)
Isotypic determinants-
Identify classes.
Are found on constant regions.
Are present among gamma globulins.
(NOT found on variable regions)
In most organ specific autoimmune diseases there is a strong association between the
disease an d HLA allele.
Immune response-
Immunogen injected in the muscle > Immunogen taken orally.
If immunogen given to a 25 yr. old > If administered to a 10 year old.
Foreign immunogen > self immunogen.
Particulate immunogen > soluble immunogen
In rheumatic fever, the possible mechanism for inducing the autoimmune phenomenon is-
B-Cell lymphoma-
Cell has DNA rearrangement detected by Southern blot using an
immunoglobulin DNA probe.
T-Cell lymphoma
Cell has DNA rearrangement detected by Southern blot using T-cell DNA
probe.
Macrophage cell line
Cell has NO rearranged DNA detected by either probe.
DO NOT Suspect a primary immunodeficiency for a 65 yr. old with recurrent infections,
total immunoglobulin levels are 35 mg/mi with a monoclonal spike by serum protein
electrophoresis.
Type B blood
ABO blood typing, serum will agglutinate blood from a type A
individual
Has anti-A antibodies in serum.
Rbc’s will agglutinate in the presence of an equivalent concentration of
anti-B antiserum. (Agglutinates with anti-B antibodies.)
NEGATIVE agglutination will look like a button of rbc’s on the bottom of the
tube.
(Serum does not have anti-B anti bodies in it.)
Type AB blood-
Has NO antibodies to ABO blood.
Will agglutinate if incubated with anti-A or anti-fl antiserum, since it has
both A and B antigens.
Anti-mumps titer of serum from A Is 1,520 and the anti-mumps titer of serum front B I:
2; the following are possible individual A has had mum~.
Individual A has been vaccinated against mumps.
Individual B has not been exposed to mumps.
Individual B has not been vaccinated against mumps.
B Lymphocytes-
Have an integral membrane Ig on their surface.
MHC Class I -
Wide cell distribution, Contains beta 2 microglobulin
Composed of 2 non-covalently linked polypeptide chains (Not of equal
molecular weight), present on most cells and almost all nucleated cells
(Not just B lymphocytes).
HLA-A, HLA-B, HLA-C (Not T1a).
The A locus encodes for Class I antigens, serologically defined by
microcytotoxicity testing.
CD8 + cytotoxic T-lymphocytes recognized antigen of MIHC I recognized by
CTL effectors.
MHC Class II Antigens
Composed of 2 non covalently linked polypeptide chains (α and β).
CD4 + helper T-lymphocytes recognize antigen as MHC II.
On surface of antigen-presenting cells that activate helper T cells.
F(ab)2 fragments-
Result from cleavage of IgG by pepsin,
Have 2 combining sites for antigen.
Fixes complement
PGE2: Pain
Neutralizing antibodies-
Largely responsible for preventing influenza virus infection.
T cell maturation –
Rearrangement of DNA which codes for T-cell receptor.
Migration to cortex to the medulla
Migration to periphery.
Division in response to stimulating antigens.
Anamnestic response is faster and more specific, longer lasting and greater then primary
immune response.
Staphylococcus aureus –
Gram + cocci,
Produces coagulase,
Catalase positive
Mannitol fermentation
Perform a catalase test, the an accustaph test
Certain strain produces an exfoliative toxin and a disease called Scaled Skin
Syndrome
Outbreak of Staph food poisoning from contaminated common food supply.
Cryptococcus neoformans –
(Not acid-fat,
Not cause chronic subacute mycosis
Not cause mycetoma, dermatomycosis, not griseofulvin D.O.C.)
Identify:
Latex Agglutination Tests- Staph aureus
Strep. pyogenes group A
Lipopolysaccharide = an Endotoxin
Endotoxin - Lipid A
Causes gram (-) sepsis
Fever and endotoxic shock
Hypotension and anoxia of organs like heart, lung and brain
Composition of cell wall determines whether a bacterial cell stains gram + or (-)
Disease: Granuloma inguinale Donovan bodies are seen from histology of genital
lesion.
Bacteroides –
Gram negative rods in anaerobic disease
Core polysaccharide –
Made of 5 sugars: Glucose, galactose, N-acetylglucosamine, heptose, and
keto-deoxyo oclulonic acid.