VISUALISATION OF INFECTION
and IMMUNE CELL ACTIVITY
J. Skopalík
7.12.2021
Typical scheme
of VIRUS attack
and replication
inthe human cell:
Virus HIV – visualization by electron microscopy
In next pages, there will be focusing and methods for micro-view
to Macrophage and Lymphocytes action
Remeber: in last lesson, we had some principial theory of immune
cell system
Bacteria capture and lysis in lysosome visualization
by electron microscopy
T lymph. / dendritic cell
How to make visualisation of IMMUNE CELL
and their INTERACTION with PATOGEN or anoter CELL ????
First method: microscopy (good for science but not for hospital daily analysis)
SURFACE MOLECULE. Antibody Ant.+ Fluorescence Probe
Second method: flow-cytometry (good also for hospital daily analysis)
Physician take the blood sample or bone marrow bioptic sample, the
cyometry is used for example to analysis of ratio CD20 cells : CD8 cells
or analysis of decreasing of CD8 lymphocyte and many other quantitative
blood and immune cell analysis….
Staining and resolution of cells is based on staining of typical surface molecules:
CD4
CD4 is a T helper cell marker, which is a single chain
transmembrane protein. The extracellular structure belongs
to IgSF, and there are four IgSF domains. The first and second
domains can bind to MHC class II molecules. CD4 acts as a coreceptor
for the TCR-CD3 complex recognition antigen and
participates in signal transduction by binding to the MHC class
II molecule, p56lek kinase.
CD8
CD8 is a cytotoxic T cell marker, a heterodimer formed by the
linkage of α and β chains by disulfide bonds, and the
extracellular structure is an IgSF member. The cytoplasmic
region of CD8 molecule can be combined with p56lek kinase
to participate in signal transduction. CD8+T lymphocytes,
which can specifically kill target cells, have anti-tumor,
antiviral and important immunomodulatory effects, and their
main function is to inhibit the immune response [15].
CD4 and CD8 molecules divide T cells into two distinct
subpopulations. CD4 and CD8 are receptors of MHC class II or
MHC class I molecules, respectively, and the changes in the
number and ratio of CD4+ and CD8+ cells reflect the immune
function status of the body.
typical surface molecules on T lymphocytes:
The cells after stining by fluorescence antibody are collected in tube and droped ONE-AFTER-ANOTHER through the light of
laser. After laser excitation, CD8 lymphocytes are mainly green fluorescence, CD4 lymphocyte are red fluorescence and
detectors (D1 and D2 on figure ) with computer compute the number of different type of cells in solution.
VACCINES
J. Skopalík
7.12.2021
• Vaccines exploit the extraordinary ability of the highly evolved human
immune system to respond to, and remember, encounters with
pathogen antigens.
• A vaccine is a biological product that can be used to safely induce an
immune response that confers protection against infection and/or
disease on subsequent exposure to a pathogen. To achieve this, the
vaccine must contain antigens that are either derived from the
pathogen or antigen produced synthetically to represent components
of the pathogen.
Statistical overview
of post-vaccination
elimination of diseases in
Britain
• The adaptive immune
response is mediated by
B cells that produce
antibodies (humoral
immunity) and by T cells
(cellular immunity). All
vaccines in routine use,
except BCG (which is believed to induce T cell
responses that prevent severe disease and innate
immune responses that may inhibit infection; see
later), are thought to mainly
confer protection
through the induction of
antibodies (Fig. 3)
Traditional
vaccines
Modern
vaccine
• Detail literature source for advance study:
A guide to vaccinology: from basic principles to new developments
Andrew J. Pollard
https://www.nature.com/articles/s41577-020-00479-7.pdf
Vaccination strategies - An overview
Giuseppe Del Giudice∗IRIS Research Center, Chiron SpA, Via Fiorentina 1,
BASIC CELL GENETICS
PEA
Mendel investigated why first
generation F1 and second
generation F2 of Pea Plant have
unlogical color heredity
J.G. Mendel - mathematician,
biologist, Augustinian friar and
abbot of St. Thomas'
Abbey in Brno
BASIC GENETIC FACTS
Human has many genes
(color of eye, gene for
hemoglobin structure, …
• Mendel’s basic conclusions for gene delivery – MENDEL LAWs
1.Law of Segregation: When gametes form, alleles are separated so
that each gamete carries only one allele for each gene
2.Law of Independent Assortment: The segregation of alleles for
one gene occurs independently to that of any other gene*
3.Law of Dominance: Recessive alleles will be masked by dominant
alleles†
* The law of independent assortment does not hold true for genes
located on the same chromosome (i.e. linked genes)
FIRST MENDEL LAW
• Simple, but not very good illustration :
• Better ilustrations:
• Why are 4 alelles at final stage stage oriniated from
one parentcell??
• Remeber MIOSIS Prophase
scheme in previous lessons :
FIRST MENDEL LAW
After chromoson
duplication
Delivery of
different T or t
into Gamets
After
ANAPHASE I
CELL before MEIOSIS
(T-gene in dominant form
t -gen in recesive form )
THIRD MENDEL LAW
(law of dominance)
•The pair of genes can be homozygous (TT or tt)
or heterozygous (Tt) , and in the case of
heterozygous pairs, one of the factors dominates
the other.
•The character that dominates ( T ) is called the
dominant character, and the one that remains
unexpressed (t ) is the recessive character.
•The recessive character, even though latent, is
transmitted to the offspring in the same way as
the dominant character.
•The recessive character is only expressed when
the offspring has two copies of the same allele
resulting in a homozygous individual.
SECOND MENDEL LAW
Mendel laws have some exception in some
organism: example where DOMINANT is not full:
…or some other exception (the case of human surface molecules of
erytrocytes)
MEMORIZE some antoher important
definiton from basic genetic:
Mendel law are used in biotechnology also for
precomputing (prediction) of possible dihybrids
(combinations of 2 genes:
Final notes to some genetic disorders:
• Not only one gene caused one function (or disfunction)
Overview: important PHENOMENON and
definiton from more advance genetic theory