NEET MDS Lessons
General Microbiology
ANTIGEN-ANTIBODY REACTIONS
I. NATURE OF ANTIGEN-ANTIBODY REACTIONS
A. Lock and Key Concept
The combining site of an antibody is located in the Fab portion of the molecule and is constructed from the hypervariable regions of the heavy and light chains. Antigen-antibody reactions is one of a key (i.e. the antigen) which fits into a lock (i.e. the antibody).
B. Non-covalent Bonds
The bonds that hold the antigen to the antibody combining site are all non-covalent in nature. These include hydrogen bonds, electrostatic bonds, Van der Waals forces and hydrophobic bonds.
C. Reversibility
Since antigen-antibody reactions occur via non-covalent bonds, they are by their nature reversible.
II. AFFINITY AND AVIDITY
A. Affinity
Antibody affinity is the strength of the reaction between a single antigenic determinant and a single combining site on the antibody. It is the sum of the attractive and repulsive forces operating between the antigenic determinant and the combining site of the antibody .
B. Avidity
Avidity is a measure of the overall strength of binding of an antigen with many antigenic determinants and multivalent antibodies. Avidity is influenced by both the valence of the antibody and the valence of the antigen. Avidity is more than the sum of the individual affinities.
III. SPECIFICITY AND CROSS REACTIVITY
A. Specificity
Specificity refers to the ability of an individual antibody combining site to react with only one antigenic determinant or the ability of a population of antibody molecules to react with only one antigen. In general, there is a high degree of specificity in antigen-antibody reactions.
B. Cross reactivity
Cross reactivity refers to the ability of an individual antibody combining site to react with more than one antigenic determinant or the ability of a population of antibody molecules to react with more than one antigen.
NON-SPECIFIC KILLER CELLS
Several different cells including NK and LAK cells, K cells, activated macrophages and eosinophils are capable of killing foreign and altered self target cells in a non-specific manner. These cells play an important role in the innate immune system.
A. NK and LAK cells
Natural killer (NK) cells are also known as large granular lymphocytes (LGL) because they resemble lymphocytes in their morphology, except that they are slightly larger and have numerous granules.
NK cells can be identified by the presence of CD56 and CD16 and a lack of CD3 cell surface markers.
NK cells are capable of killing virus-infected and malignant target cells but they are relatively inefficient in doing so.
However, upon exposure to IL-2 and IFN-gamma, NK cells become lymphokine-activated killer (LAK) cells, which are capable of killing malignant cells.
Continued exposure to IL-2 and IFN-gamma enables the LAK cells to kill transformed as well as malignant cells. LAK cell therapy is one approach for the treatment of malignancies.
NK and LAK cells have two kinds of receptors on their surface – a killer activating receptor (KAR) and a killer inhibiting receptor (KIR).
When the KAR encounters its ligand, a killer activating ligand (KAL) on the target cell the NK or LAK cells are capable of killing the target. However, if the KIR also binds to its ligand then killing is inhibited even if KAR binds to KAL.
The ligands for KIR are MHC-class I molecules. Thus, if a target cell expresses class I MHC molecules it will not be killed by NK or LAK cells even if the target also has a KAL which could bind to KAR.
Normal cells constitutively express MHC class I molecules on their surface, however, virus infected and malignant cells down regulate expression of class I MHC. Thus, NK and LAK cells selectively kill virus-infected and malignant cells while sparing normal cells.
B. K cells
Killer (K) cells are not a morphologically distinct type of cell. Rather a K cell is any cell that mediates antibody-dependent cellular cytotoxicity (ADCC).
In ADCC antibody acts as a link to bring the K cell and the target cell together to allow killing to occur. K cells have on their surface an Fc receptor for antibody and thus they can recognize, bind and kill target cells coated with antibody.
Killer cells which have Fc receptors include NK, LAK, and macrophages which have an Fc receptor for IgG antibodies and eosinophils which have an Fc receptor for IgE antibodies.
Method of Sterilization for common items
Autoclaving : Animal cages, Sugar tubes, Lab. Coats, Cotton , Filters, Instruments Culture media, Rubber, Gloves , Stopper, Tubing, Slides, Syringe and Wax needles , Test tubes, Enamel metal trays ,Wire baskets, Wood, Tongue depressor, Applicator, Endodontic instruments, Orthodontic pliers , Orthodontic kits, Saliva ejector, Handpieces Cavitron heads, Steel burs, Steel tumbler, Hand instruments
Hot air oven
Beakers, Flasks, Petri dish, Slides, Syringes, Test tubes, Glycerine, Needles ,Oil, Paper Saliva ejector, Matrix Band
Ethylene oxide
Fabric, Bedding, Blanket, Clothing, Matteresses, Pillows, Disposable instruments , Instruments, Blades, Knives, Scalpels, Scissors ,Talcum powder, Books, Cups, plates , Plastics., Flask, Petridish, Tubes, Tubing, Rubber , catheters, Drains, Gloves ,Special items - Bronchoscope, Cystoscope, Heart lung machine
Glutaraldehyde
Orthodontic kits, Orthodontic pliers , Steel burrs, 3 in 1 syringe tips ,Cystoscope ,Endoscope
Filtration
Antibiotics, Serum, Vaccines
Application of agglutination reactions
Agglutination reaction Example
Tube agglutination -> Widal test, Weil Felix reaction, Standard tube test for brucellosis
Slide agglutination -> Typing of pneumococci,Diagnosis of Salmonella,Diagnosis of Shigella
Agglutination Absorption test -> Salmonella diagnosis
Coagglutination -> Grouping of streptococci, Identification of gonococci, Detection of Haemophilus, Antigen in CSF
Passive agglutination
Latex agglutination Detection of HBs Ag, ASO, CRP
Radioimmunoassays (RIA)
It is an extremely sensitive technique in which antibody or antigen is labelled with a radioactive material. The amount of radioactive material in the antigen-antibody complex can be measured with which concentration of antigen or antibody can be assayed. After the reaction ‘free’ and ‘bound’ fractions of antigen are separated and their radioactivity-measured.
CROSS INFECTION AND STERLIZATION IN DENTISTRY
Cross infection is defined as the transmission of infectious agents amongst patients and staff with in hospital environment.
Routes of Infection
Two routes are important : transdermal and respiratory.
In transdermal route microorganisms enter the tissues of the recipient by means of injection through intact skin or mucosa (usually due to an accident involving a sharp instrument) or via defects in the skin e.g. recent cuts and abrasions.
Microorganisms causing cross infection in dentistry
Transmitted through skin
Bacteria : Treponema pallidum, Staphylococcus aureus
Viruses :Hepatitis virus, HIV ,Herpes simplex virus, Mumps, Measles , Epstein-Barr virus
Fungi: Dermatomycoses, Candidiasis,
Transmitted through aerosols
Bordetella pertussis, Myco.tuberculosis, Streptococcus pyogenes, Influenza virus
Rhinovirus, Rubella
The cell cycle
1) Labile cells (GI tract, blood cells)
- Described as parenchymal cells that are normally found in the G0 phase that can be stimulated to enter the G1
- Undergo continuous replication, and the interval between two consecutive mitoses is designated as the cell cycle
- After division, the cells enter a gap phase (G1), in which they pursue their own specialized activities
• If they continue in the cycle, after passing the restriction point (R), they are committed to a new round of division
• The G1 phase is followed by a period of nuclear DNA synthesis (S) in which all chromosomes are replicated
• The S phase is followed by a short gap phase (G2) and then by mitosis
• After each cycle, one daughter cell will become committed to differentiation, and the other will continue cycling
2) Stable cells (Hepatocytes, Kidney)
- After mitosis, the cells take up their specialized functions (G0).
- They do not re-enter the cycle unless stimulated by the loss of other cells
3) Permanent cells (neurons)
- Become terminally differentiated after mitosis and cannot re-enter the cell cycle
- Which cells do not have the ability to differentiate -> Cardiac myocytes