Homework checkQ&AAny Question from you?NTTH-HCMIU-IM-2014Any Question from you?Cells of the ISDIR-page 19-58: SUMMARY1. Cells of the IS:Who are they?Where are they?Where do they come from (Origin of immune cells )?NTTH-HCMIU-IM-2014Where do they come from (Origin of immune cells )?COMPONENT OF THE ISCELLS and SOLUBLE MEDIATORSNTTH-HCMIU-IM-2014Fig 1.1 DIRMyeloid lineageLymphoid lineageNTTH-HCMIU-IM-2014Derived from common progenitor cell in bone marrow- Pluri-potent hematopoietic stem cellINNATEADAPTIVECELLS OF INNATE IMMUNITY- Myeloid lineage1. Phagocytic cells (Phagocytes):1.1 Mononuclear phagocytes: Monocytes (circulating cells) and Macrophages (reside in tissues)1.2 Polymorphonuclear granulocytes (Polymorphs): with lobed, irregularly shaped- polymorphic- nucleusNTTH-HCMIU-IM-20142. NK cells, mast cells and plateletsCELLS OF INNATE IMMUNITY- PHAGOCYTES1.1. Monocytes/ Macrophages-Leukocytes with bean-shaped,horseshoe- shaped or brain-likeconvoluted nuclei;- relatively large (10-18 um in diameter)- Circulate in blood with half life of 8hours;- contain azurophilic granules (dyedblue)NTTH-HCMIU-IM-2014blue)- contain well- developed Golgi andmany lysosomes.- Precursors of tissue macrophagesMacrophages: Mononuclear phagocytic cells in tissue; Derive from blood monocytesHow could these cells recognize, adhere and eat the microbial and even tumor cells?- Scavenger receptors, TLRs, mannose receptors (particular sugar, lipid moleculeson microbes), FcR, C3a/ C5a R: check them laterCELLS OF INNATE IMMUNITY- PHAGOCYTES1.2. Polymorphonuclear leukocytes- Nuclei are multilobed (2 to 5); Granulocytes (Cytoplasmic granules)- Are short-lived (2-3 days), released from BM, 7 million/ min, 10- 20 um.- Based on how the granules are stained with acidic and basic dyes (HEhistological staining) they are classified into:- Neutrophil (PMN, 95% of allcirculating granulocytes): Principalphagocytic cell of innate immunity, multilobed nucleus, granules stained with bothacidic and basic dye (neutral pink)NTTH-HCMIU-IM-2014acidic and basic dye (neutral pink)- Eosinophil: Principal defender againstparasites, bi-lobed nucleus, granulesstained with acidic dye such as eosin(bright red)- Basophil: Functions similar toeosinophils and mast cells, segmentednucleus, granules stained with basic dyesuch as hematoxylin (dark blue)CELLS OF INNATE IMMUNITY- PHAGOCYTES1.2. Polymorphonuclear leukocytes (PMNs)Neutrophil (PMN, 95% of all circulating granulocytes):Principal phagocytic cell of innate immunity, multi-lobed nucleus, granules stained with both acidic and basic dye(neutral pink), 40-75 % leukocytesGranule type ProteinAcid hydrolases, muramidase,NTTH-HCMIU-IM-2014azurophilic granules (or "primary granules")myeloperoxidase, bactericidal/permeability-increasingprotein (BPI), Defensins, and the serineproteases neutrophil elastase and cathepsin Gspecific granules (or "secondary granules")Akaline phosphatase, lysozyme, NADPHoxidase, collagenase, Lactoferrin ; Cathelicidin;tertiary granules cathepsin and gelatinaseSource: Wikipedia with modificationCELLS OF INNATE IMMUNITY- PHAGOCYTES 1.2. Polymorphonuclear leukocytesEosinophil: Principal defender against parasites, bi-lobed nucleus, granules stained with acidic dye such as eosin (bright red); 1-6 % leukocytes- Granules: Major Basic Protein (MBP), Eosinophil Cationic Protein(ECP); Eosinophil-Derived Neurotoxin (EDN)- MBP: potent toxin for helminth worms, induces histamine release from mast cells, activate neutrophils and platelets and of relevance to NTTH-HCMIU-IM-2014from mast cells, activate neutrophils and platelets and of relevance to allergy, provokes bronchospasm.- Release of these molecules is the only way in which eosinophils can kill large pathogens which cannot be phagocytosed.CELLS OF INNATE IMMUNITY- PHAGOCYTES1.2. Polymorphonuclear leukocytesBasophil: Functions similar to eosinophils and mast cells i.e play a role in both parasitic infections and allergies, segmented nucleus, granules stained with basic dye such as hematoxylin (dark blue), less than 1%How? FcR; granules: histamine, proteoglycan (heparin, chondroitin), NTTH-HCMIU-IM-2014proteolytic enzymes (elastase, lysophospholipase)Mast cells?NTTH-HCMIU-IM-2014Exercise-1 WHO is WHO?The five main WBC typesNTTH-HCMIU-IM-2014CELLS OF INNATE IMMUNITY-NK, mast cells and platelets1. Phagocytic cells (Phagocytes):1.1 Mononuclear phagocytes: Monocytes (circulating cells) and Macrophages (reside in tissues)1.2 Polymorphonuclear granulocytes (Polymorphs): with lobed, irregularly shaped- polymorphic- nucleus2. NK cells, mast cells and plateletsNTTH-HCMIU-IM-2014Mast cellsNot found in circulation (i.e. tissue only, located in mucous membrane and connective tissue throughout body), very similar to basophils in their characteristics; Major effector cell in allergy; Modulation of initial immune responseNTTH-HCMIU-IM-2014NOW is yesPlatelets = cell fragments derived from megakaryocytes in the BM; 10A11 platelets/ day, 30% stored in spleen Involves in blood clot- platelet plug and fibrin clot formation (granules- alpha and delta, microtubules, actin/ myosin filaments), Alpha granules: insulin-like growth factor 1, platelet-derived growth factor, TGFp, platelet factor 4 (which is a heparin-binding chemokine) and other clotting proteins (such asthrombospondin, fibronectin, and von Willebrand factor).NTTH-HCMIU-IM-2014 Dense granules (also known as dense bodies or delta granules) are specialized secretoryorganelles: ADP, ATP, ionized calcium (which is necessary for several steps of the coagulation cascade), histamine and serotonin.WHAT PLATELETS DO....1. Stick to damaged blood vessels- requires von Willebrand factor2. Spread out to cover damaged area3. Activate and release contents- partly blocked by Aspirin4.Aggregate: Cause blood vesselNTTH-HCMIU-IM-2014constriction Cause retraction ofclot to draw wound edges togetherNK cells (natural killer)NK- Large granular lymphocytes (LGLs), ~ 15% lymphocytes, do not express BCR and TCR. Able to recognize and kill via apoptosis virus-infected cells and certain tumor cells involving activating and inhibitory receptors of NK cells (interact with MHC-I) and ALSO able to kill via ADCC mechanism.NTTH-HCMIU-IM-20141: The small lymphocyte (B, T) has no granules, round nucleus and high N:C ratio2: The LGL has azurophilic granules, indented nucleus and lower N:C ratio; CD16+CD56+ (NK cells), others (~ 5% Th, 30-50% Tc)Myeloid lineageLymphoid lineageNTTH-HCMIU-IM-2014Derived from common progenitor cell in bone marrow- Pluri-potent hematopoietic stem cellINNATEADAPTIVECELLS OF THE ADAPTIVE IMMUNITYLymphocytes: 6-10 um in diameter, 20- 50% of circulating leucocytesT cells (CD3, CD4/ CD8) and B cells (CD19)Marker molecules: communicating with environmentNTTH-HCMIU-IM-2014THE CLUSTER OF DIFFERENTIATION (CD) A protocol for identification and investigation of cell surface molecules CD number assigned on basis of 1 cell surface molecule recognized by 2 specific monoclonal antibodies CD nomenclature established in 1982NTTH-HCMIU-IM-2014 CD nomenclature established in 1982 1stInternational Workshop and Conference on Human Leukocyte Differentiation Antigens (HLDA)THE CLUSTER OF DIFFERENTIATION (CD) Examples of CD markers on leukocytesGranulocyteCD45+, CD15+Monocyte CD45+, CD14+T lymphocyteCD45+, CD3+NTTH-HCMIU-IM-2014T helper lymphocyte CD45+, CD3+, CD4+T cytotoxic lymphocyteCD45+, CD3+, CD8+B lymphocyteCD45+, CD19+Natural killer cell CD45+, CD16+, CD56+, CD3-CELLS OF THE ADAPTIVE IMMUNITYT cells:85-95 % of circulating lymphocytes T cell subsets: based on TCR and CDTh (CD4+): - MHC-IITh1 secretes IL-2;Th2 secretes IL-4, IL-5, IL-6, IL-10NTTH-HCMIU-IM-2014Th2 secretes IL-4, IL-5, IL-6, IL-10Tc (CD8+): -MHC-INKT cells: have both T cell markers andsome NK cell markersCELLS OF THE ADAPTIVE IMMUNITYB cells:5- 15%of ci r cul at i ngl ymphocyt es;BCR complex: Immunoglobulin (Ig)+ CD79a/b (Iga/b)Most express sIgM and sIgDSome of circulating B cells (~ 10%) express IgG, IgA orIgE while many more of localizing B cells express these markersNTTH-HCMIU-IM-2014B-1 cells: respond well to TI Ag (Ag stimulate B cells without T cell help), secrete natural Abs- of low avidity, but unusually they are poly reactive and are found at high concentration in the adult serum, mostly IgM but also some IgG and IgAB-2 cells (conventional B cells): respond to TD AgCELLS OF THE ADAPTIVE IMMUNITYB cells: differentiate into Ab-secreting plasma cells:B cells^ Ab-forming cells (AFCs)^ Plasma cells (infrequent in circulation, restricted in secondary lymphoid organs and BM)Plasma cells: survive for only a few daysNTTH-HCMIU-IM-2014Myeloid lineageLymphoid lineageNTTH-HCMIU-IM-2014Derived from common progenitor cell in bone marrow- Pluri-potent hematopoietic stem cellINNATEADAPTIVEAntigen presenting cellsAPCs: Antigen presenting cells- a variety of cell types thatcarry antigens in a form that can stimulate lymphocytesNTTH-HCMIU-IM-2014Who are APCs?1. Dendritic cells: Cells with dendriform (star shaped) morphology- DC (iDC) (interdigitating) found primarily in skin, lymph nodes and spleen and within or underneath most mucosal epithelia and also in thymus.from BM, interact mainly with T cellsDC1- myeloid DC- from myeloid progenitor,DC2- plasmacytoid DC- from lymphoid progenitor)NTTH-HCMIU-IM-2014DC2- plasmacytoid DC- from lymphoid progenitor)- FDC (follicular): found in the primary and secondary follicles of B cell area in lymph nodes, spleen and MALT not from BM, interact with B cells. In the B cell follicles of lymphoid organs and tissues, FDCs bind small immune complex (IC, iccosomes) and present to specific B cells.Who are APCs? DCsDifferent kinds of antigen-presenting cells (APCs)NTTH-HCMIU-IM-2014Dendritic cells- DCNTTH-HCMIU-IM-2014Who are APCs?2. Macrophages3. B cellsProfessional antigen- presenting cells: those that express MHC class II (Major Histocompatibility Complex class II ) moleculesNTTH-HCMIU-IM-2014Non- professional APCs A non-professional APC does not constitutively expressMHC class II proteins required for interaction with naive T cells; these are expressed only upon stimulation of the non- professional APC by certain cytokines such as IFN-Y. Non-professional APCs include:NTTH-HCMIU-IM-2014 Non-professional APCs include:- Fibroblasts (skin)- Thymic epithelial cells- Thyroid epithelial cells- Glial cells (brain)- Pancreatic beta cells- Vascular endothelial cellsCells of IS- Requirement Origin Characteristics Functions/ Roles and position in innate/adaptive immune responseNTTH-HCMIU-IM-2014Looking back to the previous lectureCOMPLETE BLOOD COUNT WITH DIFFERENTIAL (CBC WITH DIFF)References RangesErythrocytes (RBC) 4.0 to 5.4* M/uLThrombocytes (Platelets) 145 to 400 K/uLLeukocytes (WBC) 4.8 to 10.8 K/uLNeutrophils 40 to 74 %Band neutrophils 0 to 9 %NTTH-HCMIU-IM-2014Band neutrophils 0 to 9 %Eosinophils 0 to 6 %Basophils 0 to 1 %Lymphocytes 15 to 47 %Monocytes 0 to 12 %*: You have the chance to check this number of your own blood in the practical course by simply countingSUMMARYNTTH-HCMIU-IM-2014ORIGIN OF CELLS OF THE IMMUNE SYSTEM Derived from common progenitor cell in bone marrow Pluripotent hematopoietic stem cell Progenitor Stem Cells1. Erythroid lineageNTTH-HCMIU-IM-20141. Erythroid lineage Erythrocytes and Megakaryocytes2. Myeloid lineage Monocyte/macrophage, dendritic cells, PMNs, mast cells3. Lymphoid lineage Small and large lymphocytesCELLS OF INNATE AND ADAPTIVE IMMUNITY2.1 Myeloid Lineage- I Neutrophil Principal phagocytic cell of innate immunity Eosinophil Principal defender against parasitesNTTH-HCMIU-IM-2014 Basophil Functions similar to eosinophils and mast cells Referred to as Polymorphonuclear leukocytes (PMNs) Nuclei are multilobed (2 to 5) Granulocytes Cytoplasmic granulesCELLS OF INNATE AND ADAPTIVE IMMUNITY2.2 Myeloid lineage-II Monocytes Leukocytes with bean shaped or brain-like convoluted nuclei Circulate in blood with half life of 8 hours Precursors of tissue macrophagesNTTH-HCMIU-IM-2014 Precursors of tissue macrophages Macrophages Mononuclear phagocytic cells in tissue Derive from blood monocytes Participate in innate and adaptive immunityCELLS OF INNATE AND ADAPTIVE IMMUNITY2.3. Myeloid lineage-III Dendritic cells Cells with dendriform (star shaped) morphology Interdigitating reticular cells (synonym)NTTH-HCMIU-IM-2014 Capture and present antigens to T lymphocytes Mast cells Located in mucous membrane and connective tissue throughout body Major effector cell in allergy Modulation of initial immune responseCOMPLETE BLOOD COUNT WITH DIFFERENTIAL (CBC WITH DIFF)ReferencesRangesErythrocytes (RBC)4.0 to 5.4* M/uLThrombocytes (Platelets) 145 to 400K/uLLeukocytes (WBC)4.8 to 10.8 K/uLNeutrophils 40 to 74 % NTTH-HCMIU-IM-2014Band neutrophils 0 to 9%Eosinophils 0 to 6 %Basophils 0 to 1%Lymphocytes 15 to 47%Monocytes 0 to 12 %*: You have the chance to check this number of your own blood in the practical course by simply counting http://upload.wikimedia.org/wikipedia/commons/b/b5/Reference_ranges_for_blood_tests_-_white_blood_cells.pngNTTH-HCMIU-IM-2014Refer to CASE STUDY!CELLS OF INNATE AND ADAPTIVE IMMUNITY3. Lymphoid Lineage3.1. Large lymphocytes (large granular lymphocytes) Natural killer (NK) cells (CD16,CD56) Innate immunity to viruses and other intracellular pathogens Participate in antibody-dependent cell-mediated cytotoxicity (ADCC)NTTH-HCMIU-IM-2014 Participate in antibody-dependent cell-mediated cytotoxicity (ADCC)3.2. Small lymphocytes B cells (CD19) T cells (CD3, CD4 or CD8) Adaptive immunity Lymphocytes refers to small lymphocytes1. Cell of the ISRemember this IMAGE? Lets revise togetherNTTH-HCMIU-IM-2014Cells of IS- another illustrationFetal liver/ BMHSC/ CFU-GEMMNTTH-HCMIU-IM-2014Color Atlas of Immunology- 2003CFU-GMNKTDiscussion Topics1. Explain more clearly the question at P30-DIR,2. Methods to identify cell populations 3. Methods to isolate cell populationsNTTH-HCMIU-IM-20144. How T cell can distinguish between self and non-self Ags?5. Homework: critical thinking 1 & 2Homework checkQ&AAny Question from you?NTTH-HCMIU-IM-2014Any Question from you?Discussion Topics1. Why do warm-blooded, long-lived animals require particularly complex immune defense?- p4-DIR2. Why would removal of Ag lead to the decline in an immune response?- p14-DIRNTTH-HCMIU-IM-20143. Lets go to Critical Thinking, p18-DIR. Rewrite our discussion as a homework.And many more to explore in the DIR textbook