Leukocytes, lysosomes, LAMP – Lysosomal-associated membrane protein

1. Anti Radiation Vaccine
technology.
Dmitri Popov MD (Russia), PhD, Radiobiology
Advanced Medical Technology and Systems Inc. (Canada)
Maliev Slava , PhD, Radiobiology.
Jeffrey Jones, prof at Baylor College of Medicine. USA.

2. RADVac.
• Technical Report: Anti Radiation Vaccine technology.
• Dmitri Popov , Maliev Slava, Jeffery Jones.
• Full-text · Technical Report · Mar 2016
• Add resources
• File name: RADinVAC.pptx
DOI: 10.13140/RG.2.1.3874.0889

3. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• DNA Vaccine, Anti Radiation DNA Vaccine, Lysosomal-associated
membrane protein, Glycoprotein.

4. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• White blood cells (WBCs), also called leukocytes or leucocytes, are
the cells of the immune system that are involved in protecting the
body against both infectious disease and foreign invaders.
• All white blood cells are produced and derived from amultipotent cell
in the bone marrow known as a hematopoietic stem cell.
• Leukocytes are found throughout the body, including
theblood and lymphatic system.

5. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• All white blood cells have nuclei, which distinguishes them from the
other blood cells, the anucleated red blood cells (RBCs) and platelets.
• Types of white blood cells can be classified in standard ways. Two pairs of
broadest categories classify them either by structure
(granulocytes or agranulocytes) or by cell division lineage (myeloid cells or
lymphoid cells). These broadest categories can be further divided into the
five main types: neutrophils, eosinophils, basophils, lymphocytes,
and monocytes. These types are distinguished by their physical and
functional characteristics. Monocytes and neutrophils are phagocytic.
Further subtypes can be classified; for example, among lymphocytes, there
are B cells, T cells, and NK cells.
• https://en.wikipedia.org/wiki/White_blood_cell

6. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• MHC class I molecules are one of two primary classes of major
histocompatibility complex (MHC) molecules (the other being MHC
class II) and are found on the cell surface of all nucleated cells in the
body. Their function is to display peptide fragments of non-self
proteins from within the cell to cytotoxic T cells; this will trigger an
immediate response from the immune system against a particular
non-self antigen displayed with via help of MHC class I protein.
Because MHC class I molecules present peptides derived
from cytosolic proteins, the pathway of MHC class I presentation is
often called cytosolic or endogenous pathway.
• https://en.wikipedia.org/wiki/MHC_class_I

7. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Class I MHC molecules bind peptides generated mainly from
degradation of cytosolic proteins by the proteasome. The MHC
I:peptide complex is then inserted via endoplasmic reticulum into the
external plasma membrane of the cell. The epitope peptide is bound
on extracellular parts of the class I MHC molecule. Thus, the function
of the class I MHC is to display intracellular proteins to cytotoxic T
cells (CTLs). However, class I MHC can also present peptides
generated from exogenous proteins, in a process known as cross-
presentation.
• https://en.wikipedia.org/wiki/MHC_class_I

8. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• MHC class I molecules are heterodimers that consist of two polypeptide
chains, α and β2-microglobulin (b2m). The two chains are linked
noncovalently via interaction of b2m and the α3 domain. Only the α chain
is polymorphic and encoded by a HLA gene, while the b2m subunit is not
polymorphic and encoded by the Beta-2 microglobulin gene. The α3
domain is plasma membrane-spanning and interacts with the CD8 co-
receptor of T-cells. The α3-CD8 interaction holds the MHC I molecule in
place while the T cell receptor(TCR) on the surface of the cytotoxic T cell
binds its α1-α2 heterodimer ligand, and checks the coupled peptide for
antigenicity. The α1 and α2 domains fold to make up a groove for peptides
to bind. MHC class I molecules bind peptides that are 8-10 amino acid in
length .
• https://en.wikipedia.org/wiki/MHC_class_I

9. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• The peptides are generated mainly in the cytosol by the proteasome.
The proteasome is a macromolecule that consists of 28 subunits, of
which half affect proteolytic activity.
• The proteasome degrades intracellular proteins into small peptides
that are then released into the cytosol. The peptides have to be
translocated from the cytosol into the endoplasmic reticulum (ER) to
meet the MHC class I molecule, whose peptide-binding site is in
the lumen of the ER. They have membrane proximal Ig fold.

10. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• MHC class I molecules are loaded with peptides generated from the
degradation of ubiquitinated cytosolic proteins in proteasomes.
• As viruses induce cellular expression of viral proteins, some of these
products are tagged for degradation, with the resulting peptide
fragments entering the endoplasmic reticulum and binding to MHC I
molecules. It is in this way, the MHC class I-dependent pathway of
antigen presentation, that the virus infected cells signal T-cells that
abnormal proteins are being produced as a result of infection.

11. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• The fate of the virus-infected cell is almost always induction
of apoptosis through cell-mediated immunity, reducing the risk of infecting
neighboring cells.
• As an evolutionary response to this method of immune surveillance, many
viruses are able to down-regulate or otherwise prevent the presentation of MHC
class I molecules on the cell surface. In contrast to cytotoxic T
lymphocytes, Natural killer (NK) cells are normally inactivated upon recognizing
MHC I molecules on the surface of cells.
• Therefore, in the absence of MHC I molecules, NK cells are activated and
recognize the cell as aberrant, suggesting that it may be infected by viruses
attempting to evade immune destruction. Several human cancers also show
down-regulation of MHC I, giving transformed cells the same survival advantage
of being able to avoid normal immune surveillance designed to destroy any
infected or transformed cells.
• https://en.wikipedia.org/wiki/MHC_class_II

12. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• MHC (major histocompatibility complex) class II molecules are a family of
molecules normally found only on antigen-presenting cellssuch as dendritic
cells, mononuclear phagocytes, some endothelial cells, thymic epithelial
cells, and B cells.
• The antigens presented by class II peptides are derived from extracellular
proteins (not cytosolic as in class I); hence, the MHC class II-dependent
pathway of antigen presentation is called
the endocytic or exogenous pathway.
• Loading of a MHC class II molecule occurs by phagocytosis; extracellular
proteins are endocytosed, digested in lysosomes, and the
resulting epitopic peptide fragments are loaded onto MHC class II
molecules prior to their migration to the cell surface.
https://en.wikipedia.org/wiki/MHC_class_II

13. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Like MHC class I molecules, class II molecules are also heterodimers,
but in this case consist of two homogenous peptides, an α and β
chain, both of which are encoded in the MHC. The sub designation
α1, α2, etc. refers to separate domains within the HLA gene; each
domain is usually encoded by a different exon within the gene, and
some genes have further domains that encode leader sequences,
transmembrane sequences, etc.
• Because the antigen-binding groove of MHC class II molecules is open
at both ends while the corresponding groove on class I molecules is
closed at each end, the antigens presented by MHC class II molecules
are longer, generally between 15 and 24 amino acid residues long.

14. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
The human leukocyte antigen (HLA) system is a gene
complex encoding the major histocompatibility complex (MHC)
proteins in humans. These cell-surface proteins are responsible for the
regulation of the immune system in humans. The HLA gene complex
resides on a 3 Mbp stretch within chromosome 6p 21. HLA genes are
highly polymorphic, which means that they have many different alleles,
allowing them to fine-tune the adaptive immune system. The proteins
encoded by certain genes are also known as antigens, as a result of
their historic discovery as factors in organ transplants.

15. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• HLAs corresponding to MHC class I (A, B, and C) present peptides
from inside the cell. For example, if the cell is infected by a virus, the
HLA system brings fragments of the virus to the surface of the cell so
that the cell can be destroyed by the immune system. These peptides
are produced from digested proteins that are broken down in
the proteasomes. In general, these particular peptides are
smallpolymers, about 9 amino acids in length.

16. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Foreign antigens presented by MHC class I attract killer T-cells (also
called CD8positive- or cytotoxic T-cells) that destroy cells. MHC class I
proteins associate with β2-microglobulin, which unlike the HLA
proteins is encoded by a gene on chromosome 15.
• https://en.wikipedia.org/wiki/Human_leukocyte_antigen

17. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• HLAs corresponding to MHC class II (DP, DM, DOA, DOB, DQ, and DR)
present antigens from outside of the cell to T-lymphocytes. These
particular antigens stimulate the multiplication of T-helper cells,
which in turn stimulate antibody-producing B-cells to produce
antibodies to that specific antigen. Self-antigens are suppressed
by regulatory T cells.
• HLAs corresponding to MHC class III encode components of
the complement system.
• https://en.wikipedia.org/wiki/Human_leukocyte_antigen

18. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• HLAs have other roles. They are important in disease defense. They
are the major cause of organ transplant rejections. They may protect
against or fail to protect (if down-regulated by an infection) against
cancers. Mutations in HLA may be linked to autoimmune
disease (examples: type I diabetes, coeliac disease).
• HLA types are inherited, and some of them are connected
with autoimmune disorders and other diseases. People with certain
HLA antigens are more likely to develop certain autoimmune
diseases, such as type I diabetes, ankylosing spondylitis, celiac
disease, SLE (systemic lupus erythematosus),myasthenia
gravis, inclusion body myositis, Sjögren syndrome, and narcolepsy.

19. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• A lysosome (derived from the Greek words lysis, meaning "to loosen",
and soma, "body") is a membrane-bound cell organelle found in most
animal cells (they are absent in red blood cells).
• Structurally and chemically, they are spherical vesicles containing
hydrolytic enzymes capable of breaking down virtually all kinds
of biomolecules, including proteins, nucleic acids, carbohydrates,
lipids, and cellular debris.
• https://en.wikipedia.org/wiki/Lysosome

20. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Enzymes of the lysosomes are synthesised in the rough endoplasmic
reticulum. The enzymes are released from Golgi apparatus in small
vesicles which ultimately fuse with acidic vesicles called endosomes,
thus becoming full lysosomes. In this process, the enzymes are
specifically tagged with the molecule mannose 6-phosphate to
differentiate them from other enzymes.
• https://en.wikipedia.org/wiki/Lysosome

21. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Lysosome-associated membrane glycoproteins (lamp) are integral
membrane proteins, specific to lysosomes, and whose exact biological
function is not yet clear. Structurally, the lamp proteins consist of two
internally homologous lysosome-luminal domains separated by a
proline-rich hinge region; at the C-terminal extremity there is a
transmembrane region (TM) followed by a very short cytoplasmic tail
(C). In each of the duplicated domains, there are two
conserved disulfide bonds.

22. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• In mammals, there are two closely related types of
lamp: LAMP1 and LAMP2.
• CD69 (also called gp110 or macrosialin)is a heavily glycosylated
integral membrane protein whose structure consists of a mucin-like
domain followed by a proline-rich hinge; a single lamp-like domain; a
transmembrane region and a short cytoplasmic tail.
• CD molecules are leucocyte antigens on cell surfaces.

23. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Lysosomal-associated membrane protein 1 (LAMP-1) also known
as lysosome-associated membrane glycoprotein
1 and CD107a (Cluster of Differentiation 107a), is a protein that in
humans is encoded by the LAMP1 gene. The human LAMP1 gene is
located on the long arm (q) of chromosome 13 at region 3, band 4
(13q34).
• https://en.wikipedia.org/wiki/LAMP1

24. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Lysosomal-associated membrane protein 1 is a glyoprotein from a
family of Lysosome-associated membrane glycoproteins.[1] The LAMP-
1 glycoprotein is a type I transmembrane protein[2] which is expressed
at high or medium levels in at least 76 different normal tissue cell
types. It resides primarily across lysosomal membranes, and functions
to provide selectins with carbohydrate ligands. CD107a has also been
shown to be a marker of degranulation on lymphocytes such
as CD8+ and NK cells. And may also play a role in tumor cell
differentiation and metastasis.

25. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Although the LAMP1 glycoproteins primarily reside across lysosomal
membranes, in certain cases they can be expressed across the plasma
membrane of the cell.
• Expression of LAMP1 at the cell surface can occur due to
lysosomal fusion with the cell membrane.
• Cell surface expression of LAMP1 can serve as a ligand
for selectins and help mediate cell-cell adhesion. Accordingly, cell
surface expression of LAMP1 is seen in cells with migratory or invasive
functions, such as cytotoxic T cells, platelets and macrophages.

26. Leukocytes, lysosomes, LAMP – Lysosomal-
associated membrane protein.
• Anti Radiation DNA LUMP vaccine.
• Current therapies for Acute Radiation Syndromes and other
autoimmune diseases non-specifically suppress immune function,
• and there is great need for fundamental approaches such as antigen-
specific immune therapy.
• In this presentation we describe development of antigen-specific
immune DNA LAMP vaccines to treat Acute Radiation Syndromes in
mammals, and use of glycoprotein LAMP to enhance response to
therapy and to identify potential additional autoimmune targets for
next generation vaccines.