FISH MAJOR HISTOCOMPATIBILITY COMPLEX The MHC proteins are the cell surface glycoprotein that displays fragmented

pieces of an antigen on the host cell's surface in adaptive immunity and present processes foreign peptide fragments to specific T- lymphocytes that have the capacity to kill or co-ordinate the killing of the microbe, infected cell or malfunctioning cell in the vertebrate immune system. They may be self or nonself. If they are nonself, there are two ways by which the host cell may acquire this antigen. If the host is a leukocyte, such as a monocyte or neutrophil, it may engulf the particle (be it bacterial, viral, or particulate matter), break it apart using lysozymes, and display the fragments on Class II MHC molecules. On the other hand, if a host cell is infected by a bacterium or virus, or is cancerous, it may display the antigens on its surface with a Class I MHC molecule. Properties of MHC genes 1/ The MHC genes are highly polymorphic (nhieu dang, hnh thai) There are multi alleles of each gene, so it can make appearance of MHC molecule differ highly in individual 2/ The MHC gene is polygenic, and there are several MHC class I and MHC class II genes, encoding proteins with different ranges of peptide binding specificities. 3/ Increasing of MHC molecule The expression of both kind of MHC molecule can be stimulated by cytokines (a group of proteins and peptides that are used in organisms as signaling compounds to communicate with another in particular interferons (natural proteins produced by the cells of the immune system of most vertebrates. Interferons assist the immune response by inhibiting viral replication within other cells of the body) Structure of MHC molecule There are three main kinds of MHC molecule MHC class I MHC class II MHC class III (complement genes) All kinds of molecules belong to the immunoglobulin super family as same as T cell receptor, CD4, CD8 that all of them have antigen property themselves. Organization of MHC genes in fish Human Major Histocompatibility Complex, HLA (Human Leukocyte Antigen), This group of genes reside (c tru) on chromosome 6, and encodes cell-surface antigenpresenting proteins.

Organization of MHC class IIa and IIb genes in fish MHC class I molecule Class I molecule consist of chains attached with smaller associated chain, called 2 microglobulin. The most remarkable feature of MHC class I molecule is the structure of 1 and 2 domains which pair generate a cleft (khe) of peptide binding. MHC class I molecules are found on almost every nucleated cell of the body. Because MHC

class I molecules present peptides derived from cytosolic proteins, the pathway of MHC class I presentation is often called the cytosolic or endogenous pathway.

MHC class II molecule

MHC Class II molecules are found only on a few specialized cell types, including macrophages, dendritic cells, activated T cells, and B cells, all of which are professional antigenpresenting cells (APCs). It consists of two homologous (tng ng) peptides, and chain, both of which are encoded in the MHC. The peptides presented by class II molecules 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. The most remarkable feature of MHC class II molecule is structure of 1 and 1 domains which cooperate to generate of peptide binding.

Functions of MHC molecules MHC I As viruses infect a cell by entering its cytoplasm, MHC class I-dependent pathway of antigen presentation is the primary way for a virus-infected cell to signal T cells. It is used to presented the antigen fragments by derived(nhan c) peptides originating(khi au) in the lumen of endoplasmic reticulum within cytoplasm to the surface membrane of cell, where they are recognized by CD8 cell (cytotoxic T lymphocyte). MHC II MHC class II is to present the peptides generated in the intracellular system of B lymphocyte, magcrophage and other antigen presenting cells to CD4 cell (T helper cell). Because class II MHC is loaded with extracellular proteins, it is mainly concerned with presentation of extracellular pathogens. Class II molecules interact exclusively with CD4+ ("helper") T cells (THs). The helper T cells then help to trigger an appropriate immune response which may include localized inflammation and swelling due to recruitment of phagocytes or may lead to a full-force antibody immune response due to activation of B cells. Humoral immune response Antigen attach on the Igm of B cell and then it is engufted lysosome digest and process antigen MHC II bind to Epitope and come out to surface and recognized by CD4 on Thelper cell and TCR bind together to form a MHC antigen complex and trigger the B cell to mature B becoming plasma cell produce antibody.

Dendritic cells phagocytose exogenous pathogens, such as bacteria, parasites or toxins in the tissues . The DC uses lysosome-associated enzymes to digest pathogen-associated proteins into smaller peptides. Exogenous antigens are usually displayed on MHC Class II molecules, which interact with CD4+ helper T cells.

Cell mediated immune response

Intracellular antigens are produced by viruses replicating within a host cell. The host cell digests pathogen-associated cytoplasmic proteins by a specialized enzyme complex, the proteasome into small peptides. A specialized carrier, the transporter associated with Antigen Processing (TAP) complex moves the peptide into the endoplasmic reticulum, allowing the antigenic peptide to be coupled to an MHC Class I molecule and transported to the cell surface. MHC Class I molecules present epitope of antigen to CD8+ cytotoxic T cells. With the exception of some cell types (such as erythrocytes), Class I MHC is expressed by almost all host cells. Cytotoxic T cells (also known as TC, killer T cell, or cytotoxic T-lymphocyte (CTL)) are a population of T cells which are specialized for inducing the death of other cells. Recognition of antigenic peptides through Class I by CTLs leads to the killing of the target cell, which is infected by virus, intracytoplasmic bacterium, or are otherwise damaged or dysfunctional.

Expression of two classes of MHC molecule Lymphoid tissue T cell B cell Macrophage APC (Antigen presenting cell) Thymal epithelial cell Neutrophil Hepatocyte Kidney MHC class I +++ +++ +++ +++ + +++ + + MHC class II + +++ ++ +++ +++ _ _ _

Brain Red blood cell

+ _

_ _

Note:
An antigen-presenting cell (APC) is a cell that displays foreign antigen complexed with MHC on its surface. T-cells may recognize this complex using their T-cell receptor (TCR). There are
three main types of professional antigen-presenting cells: Dendritic cells , Macrophages , B-cells . Although almost every cell in the body is technically an APC, since it can present antigen to CD8+ T cells via MHC class I molecules, the term is often limited to those specialized cells that can prime T cells (i.e., activate a T cell that has not been exposed to antigen, termed a naive T cell). These cells generally express MHC class II as well as MHC class I molecules, and can stimulate CD4+ ("helper") T cells as well as CD8+ ("cytotoxic") T cells. Neutrophil: generally referred to as neutrophils, are the most abundant type of white blood cells and form an integral part of the immune system. Their name arrives from staining characteristics on hematoxylin and eosin (H&E) histological preparations. Neutrophils are active phagocytes, capable of ingesting microorganisms or particles. However, they can only execute one phagocytic event, expending all of their glucose reserves in an extremely vigorous "respiratory burst". The respiratory burst involves the activation of an NADPH oxidase enzyme, which produces large quantities of superoxide, a reactive oxygen species. Being highly motile, neutrophils quickly congregate(tap hp) at a focus of infection, attracted by cytokines expressed by activated endothelium, mast cells and macrophages. Hepatocyte: make up 70-80% of the cytoplasmic mass of the liver. These cells are involved in protein synthesis, protein storage and transformation of carbohydrates, synthesis of cholesterol, bile salts and phospholipids, and detoxification, modification and excretion of exogenous and endogenous substances. The expression of MHC genes in fish that the tissue distribution of MHC class I and class II mRNA After use and analyze RTPCR MHC class I high level in liver, head kidney, foregut(ruot trc)( The foregut is the anterior part of the alimentary canal, from the mouth to the duodenum(ta trang) at the entrance of the bile duct(ong mat). At this point it is continuous with the midgut. Pain in the foregut is typically referred to the epigastric(thng v) region, just below the intersection of the ribs). MHC class II express high level in kidney, spleen, hind gut, gill. Molecule markers of disease resistant

MHC I design primer on MHC I polymorphic properties, and then amplify to get DNA on that group resistant with virus. Take the fish disease and DNA extraction and amplify sequence using PCR, then compare sequences and pick areas difference and similarity and get universal primer and run PCR based isolate resistant fish. Finally, obtainning population of resistant fish from secreted bloodstock.

Antigen presentation stimulates T cells to become either "cytotoxic" CD8+ cells or "helper" CD4+ cells

In the upper pathway; foreign protein or antigen (1) is taken up by an antigen-presenting cell (2). The antigen is processed and displayed on an MHC II molecule (3), which interacts with a T helper cell (4). In the lower pathway; whole foreign proteins are bound by membrane antibodies (5) and presented to B lymphocytes (6), which process (7) and present antigen on MHC II (8) to a previously activated T helper cell (10), spurring the production of antigenspecific antibodies (9).