PH 121- Gross and Microscopic Anatomy Immune System Histology September 5, 2012 Dr.

Ryner Carrillo Lymphatic System

the second circulatory system one of the bodys primary defenses against invasion by harmful organisms and chemical toxins COMPONENTS OF THE LYMPHATIC SYSTEM Cells: -Lymphocytes of blood -Plasma cells and macrophages of connective tissue Lymphoid (a.k.a. lymphocyte-containing ) organs -Thymus -Spleen -Lymph nodes -Lymphoid nodules of the GIT ---------------------------------------------------------------4. B lymphocytesMHC MHC I -all nucleated cells -binds T cell (cytotoxic) to CD8 receptor MHC II -antigen presenting cells -binds T helper cells to CD4 receptors

Cells of the Immune System

LYMPHOCYTES Develop in the bone marrow Comprise 20-50% of WBCs in the circulation; the second most abundant WBC Most are small (~6-9m) while about 3% are large (~915m) Precursor cells develop into two types: T lymphocytes and B lymphocytes Null cells: small proportion of lymphocytes that cannot be classified as T or B lymphocytes - Capable of destroying tumor cells spontaneously or through an antibody-dependent cellular cytotoxic mechanism 1. T (thymus-dependent) LYMPHOCYTES Arise from precursor cells that migrate from the bone marrow to the thymus for further differentiation Comprise about 70% of circulating lymphocytes Antigen receptor: T cell receptor (TCR) Initial function: seek, recognize and attach to antigens that fit their surface receptors (lock and key mechanism) Ultimate function: cell-mediated immunity Functional subsets (Sensitized T lymphocytes): a. T helper cells (TH) Interact with other T and B lymphocytes to enhance the immune response help other cells to perform their effector functions by secreting a variety of mediators called interleukins Upon activation, they secrete lymphokines, which increase activation of B cells, cytotoxic T cells and suppressor T cells by antigens Attract macrophages; promote more efficient phagocytosis b. Cytotoxic T cells (TC) a.k.a. killer cells possess the ability to kill virus-infected cells and some cancer cells directly -by binding tightly to the invading cell, eventually swelling and releasing cytotoxic substances directly into the attacked cell -capable of attacking and killing many organisms in succession without being harmed interaction with TH is required for their activation and proliferation to form clones of effector cells

Figure 1. Organs of the Lymphatic System

Figure 2. WBC differentiation Leukocytes found in blood: lymphocytes, monocytes, neutrophils Soft tissues: lymphocytes, plasma cells The Story: IMMUNITY Actors: 1. APC: Macrophage, B cells, dendritic cells 2. T helper cells 3. T lymphocytes (cytotoxic lymphocytes)

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c. Suppressor T cells (TS) Inhibit the immune response -suppress the action of TH and TC -regulator of other T cells, preventing them from causing excessive immune reactions and severe tissue damage NOTE: Loss or destruction of TH (as in the case of AIDS) leads to a relative excess of TS, which therefore inhibits immune response and increases the susceptibility to infection d. Memory T cells Develop from activated T cells to provide a rapid reaction force for a subsequent encounter with the same antigen *Further classification of sensitized T lymphocytes, according to specific antigens on their membranes: a. CD4 lymphocytes - TH - account for ~70% of all T cells - activate cytotoxic cells to mature and stimulate b. CD8 lymphocytes -TS and TC -account for ~30% of all T cells A normal ratio of CD4 to CD8 lymphocytes is essential for proper immune function. (refer to the case of AIDS in the previous section) 2. B (Bone marrow-dependent) LYMPHOCYTES Derived from precursor cells in the bone marrow and continue to mature there Possess surface markers for recognition Antigen receptor: surface immunoglobulin (sIg) Initial function: synthesize and insert antibodies on their surface Ultimate function: humoral immunity Plasma cells: mature B cells that secrete antibodies (immunoglobulins: IgG, IgA, IgD, IgM, IgE) in response to antigens Memory cells: small long-lived circulating lymphocytes that are able to respond quickly to any subsequent exposure with the same antigen Figure 4. The immune response THE IMMUNE RESPONSE Initiation requires contact between antigen and surface receptors on mature lymphocytes Initiation requires contact between antigen and surface receptors on mature lymphocytes Initial phase involves lymphocyte and macrophage interaction; both are widely distributed in the body and can respond to a foreign substance wherever they encounter it. Macrophages ingest the foreign material, process its antigens and present the processed material to the lymphocytes (antigen-presenting cells, APC) Lymphocytes then respond and transform into antibodyforming plasma cells and sensitized lymphocytes, which proliferates rapidly to perform immune functions Figure 3. Antibody Antigen Presentaition 1. Antigen enters dendritic cell 2. Enzyme inside cell breaks antigen to pieces 3. Antigen pieces bind to MHC protein inside endoplasmic reticulum 4. The MHC antigen is transported to the cell surface via the Golgi apparatus 5. The MHC protein presents the antigen on the surface of the cell membrane

MAJOR HISTOCOMPATIBILITY COMPLEX (MHC) a.k.a. Human Leukocyte Antigen (HLA) set of genes that encode for the glycoproteins found on the surface of all cells Classes MHC I (self-antigen) -all nucleated cells -binds T cell (cytotoxic) CD8 receptor MHC II (T cell receptor) -Antigen-presenting cells (macrophage, dendritic cells) -binds T helper cell CD4 receptor ANTIBODY Complex protein with antigen-binding and cell-binding sites

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Following initial contact with an antigen, several weeks are needed for macrophages to process the antigen and for lymphocytes to respond After reaction to the antigen, memory cells retain the ability to respond promptly to the same antigen on subsequent exposure

Commonly associated with hypersensitivity to bacterial antigens or other foreign substances Characteristic reaction is an intense inflammatory reaction at the site of contact with the foreign substance In this process, MHC molecules of infected cells or APCs present antigenic peptides to the lymphocytes, thereby sensitizing the T lymphocytes to that antigen; this allows T lymphocytes (TH and TC) to destroy the antigen on subsequent exposure B. Humoral or Antibody Mediated Immunity

Antigen
Figure 5. Lymphocyte activation A. Cell Mediated Immunity

Recognized by sIg (surface immunoglubulin) of B-cell activates other B-cells

T-cell helper cell also recognize process antigen to release interleukins to mediate activation, clonal expansion and maturation of t-cells

Antigen taken up by APC APC - antigen presenting cells such as macrophage, B-cells, dendritic cells and Langerhans cell of the skin

clonal expansion
Some B-cells become memory cells for anamnestic response

Other B-cells mature to become plasma cells to secrete antibodies

becomes a processed antigen

binds to MHC
MHC-Ag complex contacts with a T-cell If MHC class I, it is recognized by TcR of cytotoxic T-cell
If MHC class II, TcR of helper Tcells recognize for B-cell activation antibody binds to surface antigens to block entry of organisms

plasma cells produce antibodies

antibodies eliminate antigen

binds to toxins causing inactivatio n and removal by phagocytes

antigen elimination abnormal cells recognized by cytotoxic T-cells are destroyed by apoptosis sometimes, helper T-cells release cytokines that activate macrophages that kills the organism (type IV sensitivity)

activate complimen t system

facilitates phagocytos is via opsonizatio n

antibodydependent cytotoxicity

Identification of antigen by T-cells The bodys main defense against viruses, parasites, and some bacteria Results from the function of sensitized lymphocytes Also eliminates abnormal cells that may arise during cell division (which if not destroyed, become tumors) Also causes organ transplant rejection

The major defense against many bacteria and bacterial toxins Results from the function of B lymphocytes Formation of antibodies B lymphocytes->plasma cells: response to antigen stimulation Antibodies directly attack the invader or activate the complement system, which destroys the invader In this process, macrophages present processed antigens to the lymphocytes; B lymphocyte differentiation into plasma cells follows

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Lymphatic Tissues / Organs

A. Thymus
Located in the chest, behind the sternum, and anterior to the heart FUNCTIONS: site of precursor T lymphocyte selection and maturation to T-helper and cytotoxic T lymphocytes retains non-self recognizing T cells Development of immunological selftolerance by initiating apoptosis to potentially harmful T cells Secretes thymulin, thymupoietin and thymosins to regulate T-cell maturation, proliferation and function Stem cells (precursor T lymphocytes) migrate to thymus Consist of two lobes covered by thin connective tissue and subdivided by septa into smaller lobules Should atrophy in adulthood to be replaced by adipose cells and fibrous tissue but sometimes persist STRUCTURES: 1. Capsule - beneath it is a continuous layer of epithelial reticular cells 2. Cortex (darker) - deeply staining - abundant lymphocytes - site of proliferation and selection - has a loose three-dimensional network or atypical epithelial cell called stellar reticular cell - spaces between the network of stellar reticular cell are filled with maturing lymphocytes in different stages - in the corticomedullary boundary is where macrophages are most abundant. 3. Medulla (lighter) - pale staining central portion - less lymphocytes thus epithelial cells are easier seen - maturation of self-tolerant T lymphocytes (retained cells) o Hassals Corpuscle: unique feature of medulla > eosinophilic group of flat anucleated cells > filled with keratohyalin granules > unknown significance

Figure 7. Reticular cells and Hassals Corpuscle

3 WAYS FOR NOT DEVELOPING AUTOIMMUNE T LYMPHOCYTES 1. Thymus Blood Barrier at the cortex self toxic cells contained; cannot go into circulation 2. Medullary post-capillary venules- allow selftolerant t-cells to go into circulation 3. Antigen presenting cells (APC) which present self antigen destroy autoimmune cells NOTE: Autoimmune = immune system mistakenly attacks and destroys healthy body tissue SUMMARY OF SELECTION OF T CELLS: UTERUS (has a lot of T cells but is not differentiated) FETAL LIVER PRE T CELLS THYMUS (selection of T cells) NEGATIVE SELECTION APOPTOSIS POSITIVE SELECTION CIRCULATION PERIPHERAL LYMPHOID ORGANS THYMIC BLOOD SUPPLY

Figure 6. Thymus showing lobules, medulla, and cortex

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B. Lymph Nodes
Small, encapsulated, kidney-shaped structures inserted within the pathway of lymphatic vessels Principal cell types: Lymphocytes and Reticular Cells FUNCTIONS: filters lymph drains infections as well as cancer cells distributed over the body: - prevertebral region - thorax and abdomen - mesenteries of intestines - subcutaneous CT of neck and groin Only structures in the body that have both afferent and efferent lymphatic vessels Arranged in such a way as to bring together the necessary elements for stimulation of the adaptive immune response STRUCTURES: 1. Capsule - packed collagen and reticular fibers that encloses the lymph node - has trabeculae which extend into the parenchyma - pierced by branches of afferent lymphatic vessels with valves to ensure one-way flow Subcapsular / Marginal Sinus - narrow space / sinus beneath the capsule - continuous with different sinuses: trabecular, cortical and medullary sinuses - contains macrophages, dendritic cells, lysosomes and endocytic vacuoles 2. Cortex - deeper-staining - has a greater concentration of lymphocytes Superficial Cortex - contains lymph follicles / nodules which are dense cellular aggregations with a pale-staining germinal center ; Lymph Follicles / Nodules - consistent features of the cortex of lymph nodes - site of B lymphocyte activation and proliferation - has a darker staining outer Mantle Zone / Corona which contains lymphocytes and plasma cells - pale-staining central part is the Germinal Center which is the site of development

Cortical sinuses - surrounded by dense accumulations of lymphocytes - where lymph passes through 3. Medulla - less cellular central layer of the lymph node - consists of cords of lymphoid tissue and sinuses carrying the lymph fluid Medullary Cords - longitudinal clump of cells composed of lymphocytes, macrophages, and plasma cells. - supported by reticular fibers which can only be seen using silver stain Medullary Sinuses - thin-walled structures that allow passage of lymph fluid, lymphocytes, and plasma cells out of the lymph nodes -converge to form the efferent lymphatic channels

Figure 9. Lymph Node

NOTES: o B-lymphocytes migrate to area between germinal center and subscapular sinus o T-lymphocytes migrate to inner cortex between germinal center and medulla o Activated lymphocytes migrate to medulla and differentiate into plasma cells LYMPH FLOW Afferent lymphatic vessels Subcapsular sinus Cortical sinuses which passes towards the medulla through the cortical cell mass.

Paracortical Zone - deeper regions of the cortex; consists mainly of T lymphocytes; may be greatly enlarged in a T-cell dominated response

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Medullary sinuses Hilum (concavity on one side) Efferent lymphatic vessels It then joins the blood stream via the thoracic duct or right lymphatic duct. LYMPH NODE BLOOD SUPPLY: Derived from one or more small arteries which enter at the hilum and branch in the medulla. This gives rise to extensive capillary networks corresponding to the cortical follicles, paracortical zone and medullary cords. Lymphocytes enter lymph nodes mainly via the arterial system, gaining access by migrating across the walls of specialized postcapillary venules (high endothelial venules (HEV) The HEV drain into small veins that leave the node via the hilum.
Figure 11. Distribution of lymph nodes in the body

------------------------------------------------------------NICE TO KNOW:
B Lymphocytes and other cells (Germinal Center) Resting B cells - Exposed to antigen to which they would react and enter cycle of blast transformation Centroblasts - Large, mitotically active cells with round nuclei that are found in the darker zone of the germinal center. (differentiates into centrocytes) - Undergo increased mutation of the immunoglobulin genes (somatic hypermutation), thus creating further variations in immunoglobulin structure - Differentiate into plasma cells or memory cells. Centrocytes - Found in the pale midzone of the germinal center - Are of variable size; have folded, irregular (cleaved) nuclei. Mitotic figures are absent in this area. - Migrate towards the paler capsular zone of the germinal center where they go through further division to produce either immunoblasts I or memory B cells Immunoblasts - Move to the medullary cords where they complete their differentiation into plasma cells capable of secreting large amounts of antibody. Follicular Dendritic Cells - Major antigen presenting cells of the follicles. - Found in all areas of the germinal center and also form a meshwork in the mantle zone and in primary follicles. - Can retain antigen on their surface for many months and may have a role in maintaining the activity of memory cells as well as stimulating a primary immune response. Tingible Body Macrophages - Easily seen in routine sections in active germinal centers. - Contain within their cytoplasm numerous apoptotic bodies derived from B lymphocytes that have not been successful in generating a high affinity antibody. T Lymphocytes (Paracortical Zone) - Enlarge to form immunoblasts Interdigitating Dendritic Cell - Cytoplasmic processes form a meshwork in the paracortex and are in close contact with the naive T cells circulating through this zone. - Derived from macrophage precursors including the Langerhans cells of the skin. **Paracortical reaction great expansion of the paracortical zone in a T cell-dominated immunological response **Addresins lymphocyte binding molecules that allow lymphocyte to bind to the endothelium as the first step of migration into the tissue.

Figure 10. Blood supply of lymph node

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C. Mucosa-associated Lymphoid Tissue (MALT)

aggregates of lymphoid tissue found in the digestive and reproductive systems Tonsils (palatine, lingual, pharyngeal) Adenoids Small Intestines: aggregate lymphoid nodules which were known as Peyers Patches Appendix Vagina composed mostly T lymphocytes but also have B cells and plasma cells. functions as lymph nodes lymphocytes exposed in this regions go through regional lymph nodes then back to the region after activation secretes immunoglobulins: IgA - protect against pathogens that may gain access to underlying tissues IgG- majority of antibody-based immunity against invading pathogens IgM- eliminates pathogens in the early stages of B cell mediated immunity before there is sufficient IgG - secreted into lamina propria

Removal of particulate matter and aged or defective blood cells, particularly erythrocytes, from the circulation. Haemopoiesis in the normal fetus and in adults with certain diseases; as well as storage of 1/3 the bodys platelets and as reservoir of mature RBCs

STRUCTURES: 1. Capsule - fibroelastic outer layer - forms trabeculae that extend into the parenchyma - thickened at the hilus and is continuous with supporting tissues that sheath the larger blood vessels entering and leaving the organ. 2. Splenic Parenchyma - not subdivided into a cortex and medulla - consists of red and white islands of cells Red Pulp cords (cords of Bilroth) of reticular cells and splenic sinuses - highly vascular tissue that forms the bulk of the organ - sinuses are abundant in RBCs White Pulp discrete 0.5 - 1mm white nodules - consists of B and T cells which make up 5-20% of the total mass of the spleen. - consist of lymphoid tissue; scattered through red pulp Perilymphoid/Perifollicular Zone - zone of the red pulp immediately surrounding the white pulp. -devoid of sinuses; sparse reticulin meshwork and contains a large number of red and white blood cells in the same proportion as that of blood. Mantle Zone narrow zone of small lymphocytes at the follicular periphery. Marginal Zone - broader zone of less densely packed medium-sized lymphocytes supported by a framework of reticulin fibers. Central Artery - surrounded by a periarterial sheath of diffuse and nodular lymphoid tissue

D. Tonsils
PALATINE TONSILS - stratified squamous epithelium due to abrasion - tonsil crypts

Figure 12. Tonsil with SS Epithelium and Crypt

E. Spleen
largest lymphoid organ in the body situated in the left upper quarter of the abdomen between fundus and diaphragm Receives a rich blood supply via a single artery, the splenic artery, and is drained by the splenic vein into the hepatic portal system Covered by the visceral peritoneum, which consists of a single layer of squamous epithelium FUNCTIONS: Production of immunological responses against blood borne antigens

Figure 13. Splenic parenchyma

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SPLENIC CIRCULATION Blood enters the spleen in the splenic artery, which branches within the parenchyma. The larger arteries are surrounded by central arteries surrounded by periarteriolar lymphoid sheath PALS which consists mainly of TH cell The central artery branch at right angles to form penicilliary arteries PA, and these terminate in two to three sheathed capillaries SC. These vessels do not have endothelial lining but surrounded by macrophages. The blood arriving from a sheathed capillary traverses the SC before entering the red pulp RP. The sheathed capillaries, form the first part of the filtering mechanism of the spleen. NOTE: There are two theories regarding the termination of blood vessels CLOSED CIRCULATION - the blood vessels entering the red pulp become confluent with the sinuses of the red pulp OPEN CIRCULATION (favored) - the blood vessels open at their ends, into the extracellular space of the parenchyma, permitting the extravasation of RBCs - blood goes back to circulation by going through clefts in the walls of the sinuses SPLENIC LYMPHOID TISSUE - Lymphoid tissue (T and B cells) is situated in the white pulp. The NFA (non-filtering area) of the red pulp is considered to be part of the splenic lymphoid tissue, but its immunological function remains to be unclear. T cells mass of these cells form eccentric cylindrical sheath around the central artery. (mainly helper T cells) B cells form follicles at the vicinity of an arteriole which exhibits germinal center with the function similar to the ones in the lymph nodes

END
References: Sir Carrillos Lecture Bloom and Fawcett NOTE: Bloom and Fawcett daw ang main reference ni Sir. ------------------------------------------------------------------------------------

Lab Checklist (From Sir): 1. Lymph Node a. Regions and Zones b. Structures c. Circulation 2. MALT 3. Thymus a. Regions b. Structures c. Circulation 4. Spleen a. Regions b. Structures c. Circulation
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From the sinuses, blood drains to short pulp veins Trabecular veins Splenic vein

Laughing lowers

levels of stress hormones and strengthens the immune system. Six-year-olds laugh an average of 300 times a day. Adults only laugh 15 to 100 times a day. Isang anat exam na lang after this!!!!! Good luck sa combo natin this coming Monday and Tuesday, 2014! - Team 7

Figure 14. Visualization of splenic closed and open circulation

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