ch27 the immune system
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Ch27 The Immune System. Yuki Juan NTU May 26, 2003. Outline. Introduction Antibody-based or humoral immunity Activation of B cells to produce antibodies T cells and cell-mediated immunity Why does the human immune system so fiercely reject foreign human cells Monoclonal antibodies. - PowerPoint PPT PresentationTRANSCRIPT
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Ch27 The Immune SystemCh27 The Immune System
Yuki JuanYuki Juan
NTUNTU
May 26, 2003 May 26, 2003
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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TermsTerms
Autoimmune reactions: Autoimmune reactions: – The immune system would attack the The immune system would attack the
components of the body components of the body
Autoimmune diseaseAutoimmune disease
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The Cells Involved In The Immune The Cells Involved In The Immune SystemSystem
B cells: B cells: – Multiply and undergo their primary maturation Multiply and undergo their primary maturation
in bone marrowin bone marrow
T cells: T cells: – Multiple and mature in thymus glandsMultiple and mature in thymus glands– Helper T cells and cytotoxic killer T cellsHelper T cells and cytotoxic killer T cells
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HemopoiesisHemopoiesis
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Two Immune Protective SystemsTwo Immune Protective Systems
Humoral immunityHumoral immunity
Cell-mediated immunityCell-mediated immunity
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Humoral ImmunityHumoral Immunity
Antibody: Antibody: – Combine with the foreign antigensCombine with the foreign antigens
Antigen: Antigen: – AntiAntibody body gengenerationeration
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Cell-mediated ImmunityCell-mediated Immunity
Cytotoxic T cells or killer T cells recognize Cytotoxic T cells or killer T cells recognize abnormal cells in the body.abnormal cells in the body.
The killer cell attaches to the virus-infected The killer cell attaches to the virus-infected cell and destroys it.cell and destroys it.
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The Differences Between Humoral The Differences Between Humoral And Cell-mediated ImmunityAnd Cell-mediated Immunity
The humoral system:The humoral system:– is part of the defence against virus in the bloois part of the defence against virus in the bloo
d or in the mucous membrane before it infects d or in the mucous membrane before it infects a cell.a cell.
Cell-mediated immunityCell-mediated immunity– Destroys the host cell after the latter has becoDestroys the host cell after the latter has beco
me infected and in doing so aborts virus replicme infected and in doing so aborts virus replication in that cell.ation in that cell.
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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AntibodiesAntibodies
Immunoglobulins (Igs)Immunoglobulins (Igs)
Y-shaped proteinY-shaped protein– Two identical light polypeptide chainsTwo identical light polypeptide chains– Two identical heavy chainsTwo identical heavy chains
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Structure of Antibody Structure of Antibody
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TermsTerms
EpitopeEpitope– The specific part of the antigen which is recogThe specific part of the antigen which is recog
nized by an antibodynized by an antibody
CloneClone– A collection of identical cells arising from multiA collection of identical cells arising from multi
plication of a single cellplication of a single cell
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Different Classes of AntibodiesDifferent Classes of Antibodies
IgMIgM– When the body responds to an antigen, the first produWhen the body responds to an antigen, the first produ
ced antibodyced antibody– Multi-subunit or polymericMulti-subunit or polymeric– Form of antibody with 10 antigen combining sitesForm of antibody with 10 antigen combining sites
IgGIgG– More prolonged challenge by the same antigenMore prolonged challenge by the same antigen
IgAIgA– Of importance as a first of defenceOf importance as a first of defence
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Generation Of Antiboy DiversityGeneration Of Antiboy Diversity
300 V gene fragments300 V gene fragments
4 J gene segments4 J gene segments
C gene segmentC gene segment
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The Process of Rearrangment LeadThe Process of Rearrangment Leading to a Functional L Chain Immunoing to a Functional L Chain Immuno
globulin Geneglobulin Gene
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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The Principle of Clonal SelectionThe Principle of Clonal Selection
Macfarlane Macfarlane BurnetBurnet
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Conversion of A Conversion of A Virgin B Cell to An Virgin B Cell to An Activated State, Activated State, But not yet to an But not yet to an
Antibody-Antibody-producing Plasma producing Plasma
Cell Cell MHC classes II MHC classes II proteins: on the proteins: on the surface of the B cellssurface of the B cells
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Comparison of Receptors on B and Comparison of Receptors on B and T CellsT Cells
B Cell receptorsB Cell receptors T Cell receptorsT Cell receptors
Are formed from antibodies Are formed from antibodies displayed in the cell membranedisplayed in the cell membrane
Special receptor proteins in the Special receptor proteins in the membranemembrane
Specific receptors for each cell Specific receptors for each cell generated randomlygenerated randomly
Specific receptors for each cell geSpecific receptors for each cell generated randomlynerated randomly
Bind to antigen molecules in Bind to antigen molecules in solution. Receptor recognizes the solution. Receptor recognizes the antigen itselfantigen itself
Cannot bind to antigens in solution.Cannot bind to antigens in solution. Antigen must be internalized by s Antigen must be internalized by special APC which degrades it and pecial APC which degrades it and displays peptides in MHC proteins. displays peptides in MHC proteins. The receptor recognises the complThe receptor recognises the complex of peptide-MGC protein on the ex of peptide-MGC protein on the APC, not either component aloneAPC, not either component alone
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Activation of a Helper T Cell By An Activation of a Helper T Cell By An Antigen-presenting Cell (APC)Antigen-presenting Cell (APC)
APC: A special phagocytic white cell
Cell surface markers known as clusters of differentiation
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A Helper T Cell Activating a B Cell A Helper T Cell Activating a B Cell to Become A Clone of Plasma Cells to Become A Clone of Plasma Cells
Secreting AntibodySecreting Antibody
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Summary Summary Scheme of Scheme of Antibody Antibody
Production Production by B Cellsby B Cells
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TermTerm
Memory cellsMemory cells– They are the basis of long-term immunity from They are the basis of long-term immunity from
a repeat infectiona repeat infection
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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Class II MHCs and Class I MHCsClass II MHCs and Class I MHCs
Helper T cells Helper T cells – recognize displayed peptides on class II MHCrecognize displayed peptides on class II MHC
ss– Have CD4 proteinsHave CD4 proteins
Killer T cells Killer T cells – require display on class I MHCsrequire display on class I MHCs– Have CD8 proteinsHave CD8 proteins
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Sequence of Events Sequence of Events In Cell-mediated In Cell-mediated
Immune Reaction to a Immune Reaction to a Foreign Antigen Foreign Antigen
Synthesized Inside Synthesized Inside CellsCells
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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Why Does the Human Immune SystWhy Does the Human Immune System So Fiercely Reject Foreign Humem So Fiercely Reject Foreign Hum
an Cellsan Cells
The main cause of rejection of foreign The main cause of rejection of foreign tissue grafts in the MHC molecules on tissue grafts in the MHC molecules on cells. cells.
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OutlineOutline
IntroductionIntroduction
Antibody-based or humoral immunityAntibody-based or humoral immunity
Activation of B cells to produce antibodiesActivation of B cells to produce antibodies
T cells and cell-mediated immunityT cells and cell-mediated immunity
Why does the human immune system so fiWhy does the human immune system so fiercely reject foreign human cellsercely reject foreign human cells
Monoclonal antibodiesMonoclonal antibodies
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Monoclonal AntibodiesMonoclonal Antibodies
The achievement of this earned Cesar The achievement of this earned Cesar Milstein and Georges Kohler in Cambridge Milstein and Georges Kohler in Cambridge a Nobel Prizea Nobel Prize
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單株抗體單株抗體由單一細胞株所製造由單一細胞株所製造出來的抗體出來的抗體
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多株抗體多株抗體抗血清是多株抗體抗血清是多株抗體 , , 這些抗體是由各種不這些抗體是由各種不同種同種 BB 細胞所產生抗細胞所產生抗體之混合物體之混合物
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單株抗體的特株性質單株抗體的特株性質它只和抗原中的某種抗原決定位置結合它只和抗原中的某種抗原決定位置結合 ,, 不不會與不同之抗原決定位置相結合會與不同之抗原決定位置相結合 ,, 所以有很所以有很高的專一性高的專一性它和抗原決定位置結合的強度及性質完全它和抗原決定位置結合的強度及性質完全相同相同 ,, 所以容易用它來純化抗原所以容易用它來純化抗原
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單株抗體的製造方法單株抗體的製造方法正常細胞正常細胞藉由轉型藉由轉型 (transform)(transform) 變成變成癌細胞癌細胞 ;;也就是使正常細胞表現出致癌基因也就是使正常細胞表現出致癌基因 (oncoge(oncogene)ne)
如何使抗體分泌細胞變成癌細胞如何使抗體分泌細胞變成癌細胞– 用用致癌物質致癌物質或或病毒病毒使此類細胞內表現致癌基因使此類細胞內表現致癌基因– 細胞融合技術細胞融合技術
致癌細胞之致癌基因融合到抗體產分泌細胞中致癌細胞之致癌基因融合到抗體產分泌細胞中融合融合瘤細胞瘤細胞 (hybridoma)(hybridoma)
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融合瘤細胞融合瘤細胞 (Hybridoma)(Hybridoma) 之製造之製造1975, Milstein and Kohler1975, Milstein and Kohler 發表發表 1984,1984, 諾諾貝爾生理醫學獎貝爾生理醫學獎老鼠的融合瘤細胞老鼠的融合瘤細胞脾臟脾臟 BB 細胞 細胞 + + 骨髓瘤細胞骨髓瘤細胞 以特殊溶液處理,分離融合細胞以特殊溶液處理,分離融合細胞 篩選具有特殊抗體的細胞株篩選具有特殊抗體的細胞株 自該細胞的組織培養中,收集所需抗體自該細胞的組織培養中,收集所需抗體
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單株抗體的應用單株抗體的應用檢驗試劑的開發製造檢驗試劑的開發製造醫學治療醫學治療生技產品之純化生技產品之純化農業及畜牧業農業及畜牧業基礎研究基礎研究
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單株抗體的應用單株抗體的應用 --檢驗試劑的開發製造檢驗試劑的開發製造
荷爾蒙荷爾蒙組織及細胞抗原組織及細胞抗原藥物監控藥物監控藥物濫用藥物濫用微生物微生物抗體及過敏原抗體及過敏原酵素酵素生長因子生長因子
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單株抗體的應用單株抗體的應用 --醫學治療醫學治療
器官移植器官移植腫瘤影像腫瘤影像抗癌藥抗癌藥
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Ch 29 Gene Cloning, Ch 29 Gene Cloning, Recombinant DNA Technology, Recombinant DNA Technology,
Genetic EngineeringGenetic Engineering
Yuki JuanYuki Juan
NTUNTU
May 27, 2003May 27, 2003
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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DNases and Restriction EnzymesDNases and Restriction Enzymes
DNasesDNases– The digestive enzymes of pancreatic juiceThe digestive enzymes of pancreatic juice– Cut the phosphodiester bondsCut the phosphodiester bonds– Cut DNA randomlyCut DNA randomly
Restriction enzymesRestriction enzymes– Don’t cut DNA randomlyDon’t cut DNA randomly– Each recognizes a specific short sequence of Each recognizes a specific short sequence of
bases so as to make a cut at a precision point bases so as to make a cut at a precision point in the sequence in the sequence
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基因選殖基因選殖– 1970, Smith1970, Smith 發現第二類核酸發現第二類核酸
限切酵素限切酵素,,可以很準確分割可以很準確分割 DDNANA
– 1973, Boyer-Cohen-Chang1973, Boyer-Cohen-Chang完成第一基因選殖的工作完成第一基因選殖的工作
The first to be isolated from E. coli strain R
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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CloneClone
Refers to multiple identical copies produceRefers to multiple identical copies produced from a single origind from a single origin
Genomic cloning and cDNA cloningGenomic cloning and cDNA cloning
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寄主細胞寄主細胞蛋白質表現系統主要有蛋白質表現系統主要有– 大腸桿菌大腸桿菌 : : 操作方便、便宜操作方便、便宜
缺點缺點 : : – 沒有修飾蛋白質的能力沒有修飾蛋白質的能力– 無法正確摺疊 無法正確摺疊 – 會產生毒素會產生毒素
– 酵母菌酵母菌– 昆蟲細胞昆蟲細胞 : : 操作較複雜、成本高操作較複雜、成本高– 哺乳類細胞哺乳類細胞 : : 操作較複雜、成本操作較複雜、成本
高高
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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第二類核酸限切酵素第二類核酸限切酵素
鈍端 (blunt end)黏端 (sticky end)
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連接酵素連接酵素 (ligase)(ligase)
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Sequence of Steps In Producing in Sequence of Steps In Producing in Bacteriophage a Recombinant DBacteriophage a Recombinant D
NA LibraryNA Library
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Illustration of Gene CIllustration of Gene Cloning Using Bacterioloning Using Bacteriophage Lambda Phagphage Lambda Phage as The Cloning Vee as The Cloning Ve
hiclehicle
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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cDNAcDNA
Complementary DNA: the double-stranded Complementary DNA: the double-stranded DNA copy of mRNADNA copy of mRNA
Genomic DNA: contains intronsGenomic DNA: contains introns
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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Transferring a Transferring a DNA Insert Into A DNA Insert Into A PlasmidPlasmid
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The Structures of dATP and ddATPThe Structures of dATP and ddATP
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An Autoradiograph Sequencing GelAn Autoradiograph Sequencing Gel
Dideoxy DNDideoxy DNA-sequenciA-sequencing Techniqng Techniqueue
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Photograph of an Photograph of an Actual Actual
Sequencing Sequencing LadderLadder
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DNADNA 定序定序Maxam-Gilbert MethodMaxam-Gilbert Method
G ReactionG Reaction– ATTGACTTAGCCATTGACTTAGCC– ATTGACTTAATTGACTTA– ATTATT
A and G ReactionA and G Reaction– ATTGACTTAGCCATTGACTTAGCC– ATTGACTTAATTGACTTA– ATTGACTTATTGACTT– ATTGATTG– ATTATT
T and C ReactionT and C Reaction– ATTGACTTAGCCATTGACTTAGCC– ATTGACTTAGCATTGACTTAGC– ATTGACTTAGATTGACTTAG– ATTGACTATTGACT– ATTGACATTGAC– ATTGAATTGA– ATAT– AA
C ReactionC Reaction– ATTGACTTAGCCATTGACTTAGCC– ATTGACTTAGCATTGACTTAGC– ATTGACTTAGATTGACTTAG– ATTGAATTGA
CCGATTCAGTT
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The Basic Principle Underlying The Basic Principle Underlying Amplification of A DNA Section by Amplification of A DNA Section by
PCRPCR
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聚合酵素連鎖反應聚合酵素連鎖反應
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聚合酵素連鎖反應聚合酵素連鎖反應 (1)(1)
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聚合酵素連鎖反應聚合酵素連鎖反應 (2)(2)
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聚合酵素連鎖反應聚合酵素連鎖反應 (3)(3)
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TermsTerms
Transgenesis:Transgenesis:– Genes can be inserted into animals and plantGenes can be inserted into animals and plant
s to give different phenotypess to give different phenotypes
Gene therapyGene therapy– Involves correction of a gene deficiency by insInvolves correction of a gene deficiency by ins
erting a normal gene into cellserting a normal gene into cells
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OutlineOutline
The first step-cutting the DNA with restrictiThe first step-cutting the DNA with restriction endonucleaseson endonucleases
Gene cloning, or how genes are isolatedGene cloning, or how genes are isolated
Isolation of a genomic clone of a human gIsolation of a genomic clone of a human geneene
Cloning a human cDNACloning a human cDNA
What can be done with the cloned DNAWhat can be done with the cloned DNA
Knockout mice of gene targetingKnockout mice of gene targeting
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TermsTerms
Knockout miceKnockout mice– Mice in which a specific gene has been functionally eliMice in which a specific gene has been functionally eli
minated minated
Stem cellsStem cells– Cells can divide without limitCells can divide without limit
Embryonic stem cellsEmbryonic stem cells– Different from other stem cells and differentiated cellsDifferent from other stem cells and differentiated cells– They give rise every type of cell in the bodyThey give rise every type of cell in the body– TotipotentTotipotent
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Procedure For Obtaining Knockout Procedure For Obtaining Knockout Mice By Gene-targeted MutationMice By Gene-targeted Mutation
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One Method of Gene Targeting by One Method of Gene Targeting by Homolgous Recombination in ES CHomolgous Recombination in ES C
ellsells