1. The Major Histocompatibility Complex Antigen Presentation and Processing and the Relationship to Susceptibility and Resistance to Diseases; Folder Title: MHC Updated: November 25, 2012

2. Kuby Immunology, 6 th Edition, Chapter 8 Topics Listed vs Topics Covered in BIO 447, See Page 189 1.General Organization and Inheritance of the MHC Genes 2.MHC Molecules and Genes 3.Detailed Genomic Map of MHC Genes (Not Covered in BIO 447) 4.Cellular Expression of MHC Molecules 5.Regulation of MHC Expression (Not Covered in BIO 447) 6.MHC and Disease Susceptibility 7.MHC and Immune Responsiveness 8.Self-MHC Restriction of T-Cells 9.Role of Antigen-Presenting Cells Not Covered in BIO 447 10. Evidence of Different Antigen-processing and Presentation Pathways 11. Endogenous Antigen: The Cytosolic Pathway 12. Exogenous Antigens: The Endocytic Pathway 13. Cross Presentation of Exogenous Antigens 14. Presentation of Non-Peptide Antigens

3. What we saw earlier in the course: What Affects Host Response to Potential Antigens?

4. What else affects host response to potential antigens?

5. What are we looking at? Groups of Mice with Same Collection of MHC Alleles (Haplotype) How do these groups respond to two different experimental antigens?

14. Haplotype = Combination of MHC Genes Inherited from Each Parent Co-dominantly Expressed = both sets of alleles are expressed

15. On a scale of 1 to 5: 1 = I’m totally lost; 2 I’m having a hard time but I get some of it. 3 = I’m doing OK. I get a lot of it. I’ll figure the rest out later. 4 = I’m doing fine. I get most of it; 5 = +2 = This is no problem. Please get moving before I get bored

16. Both parents are of course histocompatible with their own tissues, and their progeny tolerate skin grafts from either parent because the progeny recognizes antigens from either parent as self. Transplanting hematopoietic stem cells from parent to mixed progeny could generate graft-vs-host reaction

17. What happens if we transplant skin from one b/k progeny hybrid to another b/k hybrid? What happens if we transplant from a b/k hybrid back to either of the parental strains (b/b or k/k)?

19. Why is this collection of genes controlling whether one can transplant a tissue from one mouse to another or from one human to another? Nature doesn’t do transplants. Why does Nature bother to generate genes that control transplantability of tissue?

20. What are these genes controlling tissue compatability (“histocompatibility”)? “Histocompatability Complex or Major Histocompatability Genes = MHC Genes” What products are these genes coding for that control histocompatibility?

21. Figure 8-1, Kuby 6 th Edition, Top Half Human HLA (Histocompatibility Complex), Lower Half Next Slide Mouse H2 Complex Complement Proteins Some Cytokines

22. In Humans the Genes Corresponding to MHC Region in Mice are Called Human Leukocyte Associated Antigens or HLA Genes

23. Kuby, 6 th Edition, Figure 8.01, Bottom Half Class II Loci Equivalent to IAa, IAb; IEa, IEb in Mice Class I Loci Equivalent to K, D, and L in Mice Human HLA Loci

24. Recombinant Collection of HLA Genes not existing in this grouping in either parent

26. k and b Haplotype “Team Members” See Table 7-1, Kuby 4th Edition, p. 175

27. Haplotype and Team Members

28. These H2 Genes Control Recognition of Self when cells from mouse strains with two different H2 Haplotypes are mixed together in cell culture. Mixed Lymphocyte Test in Cell Culture

29. MLR Assay See Figure 14-16, p.367, Kuby, 6th Edition Strain X Cells are activated to proliferate by exposure to unmatched cells from Strain Y

30. Strain X Cells are also activated by exposure to strain Y cells in the living mouse Strain X mouse makes T-cells that will lyse strain Y cells

31. CML Assay See Figure 8-15, p. 207, Kuby, 6th Edition For MHC Restriction. MHC Restriction will be covered later See Figure 14-17, Kuby 6 th Edition, p. 368. For CML Assay Cell-Mediated Lympholysis Assay (CML Assay) Strain X Cells are also activated by exposure to strain Y cells in the living mouse Strain X mouse makes T-cells that will lyse strain Y cells. Why is Strain X mouse doing this? What does the strain X mouse “think” it sees? Get lysis of Strain Y target cells

32. CML Assay See Figure 8-15, p. 207, Kuby, 6th Edition For MHC Restriction. MHC Restriction will be covered later See Figure 14-17, Kuby 6 th Edition, p. 368. For CML Assay

33. Restriction to Self MHC Alleles In Vitro exposure of target cells to spleen cells from LCM virus-infected mouse.

34. On a scale of 1 to 5: 1 = I’m totally lost; 2 I’m having a hard time but I get some of it. 3 = I’m doing OK. I get a lot of it. I’ll figure the rest out later. 4 = I’m doing fine. I get most of it; 5 = +2 = This is no problem. Please get moving before I get bored

35. What Do MHC Class I and Class II Proteins Look Like? How do they bind and present peptide antigens?

37. MHC-I See figure 8-3, Kuby, 6th Edition, p. 194, left

38. MHC-II See figure 8-3, Kuby, 6th Edition, p. 194, right

39. Class I MHC Protein Binding Cleft

40. Ag Site Class I See figure 8-4, Kuby, 6th Edition, p. 195, right

41. Ag-Bind See figure 8-9(a,b), Kuby, 6th Edition, p. 199

43. Ag in Site Class I See figure 8-9(c), Kuby, 6th Edition, p. 199

44. Class I MHC Space-Filling

45. H2K and Peptides See figure 7-12, Kuby 4th edition, p. 184

46. Class I and Class II proteins have sequence differences inherited in the germ line (they are polymorphic) Where are the variations located in the Class I and Class II proteins?

47. Amino Acids Positions of Polymorphic Variations in Class I MHC Proteins in Humans Membrane Distal a1 and a2 Domains (Comprise Peptide-binding Cleft) Trans- Membrane Residues Kuby, 6th Edition, Figure 8-10(a) p. 201; MHCIVary

48. MHC-Class I Polymorphisms Kuby, 6th Edition Figure 8-10(b) p. 201 MHCIPoly Positions of Polymorphic Amino Acid Residues in a Human Class I MHC Protein

49. MHC ClassI and Flu

50. On a scale of 1 to 5: 1 = I’m totally lost; 2 I’m having a hard time but I get some of it. 3 = I’m doing OK. I get a lot of it. I’ll figure the rest out later. 4 = I’m doing fine. I get most of it; 5 = +2 = This is no problem. Please get moving before I get bored

51. MHC-II See Figure 8-3, Right Kuby, 6h Edition p. 194

52. Class II MHC Space Filling

53. HMC-Class II Dimers

54. Comparison of Class I and Class II

55. TCR and MHC-Class II

56. MHC-Class II and Bound Antigen

57. Class I and Class II Antigen Acquisition

58. See Figure 8-12, p. 203 Kuby, 6 th Edition) Variety of MHC Loci & Allelles

60. Why are there so many different Class I and Class II proteins in a given individual and in a species gene pool? Why are these gene products so polymorphic?

61. Polymorphism of Class I and Class II Molecules in Human HLA Humans: 6 Different Class I Molecules HLA-A, HLA-B, HLA-C (3 Class I from one parent, 3 from the other) Approximate HLA Class I Alleles in Persons of European Descent: HLA-A: 60 HLA-B: 110HLA-C: 40 Humans: 12 Different Class II Molecules (Hetero-dimers) DPa,DPb, DQa,DQb, DRa, DRb (6 Class II from one parent, 6 from the other) Approximate HLA Class II Alleles in Persons of European Descent: 122 Alleles of HLA-DRbeta alone Theoretical Combination of possible alleles is virtually infinite Combination is inherited at birth Does not diversify in an individual the way TCR or Antibodies do. Diversity among individuals depends on degree of familial relationship (full siblings histocompatible about 25% of the time) Linkage Disequilibrium favors allele combinations based on (1) time of divergence from population founder, (2) hot-spots facilitating genetic cross-overs, (3) selection for or against certain combinations MHCAlleles

62. Association of Human MHC Alleles and Risk for Diseases (from Table 7-4, Kuby Immunology, 4th Edition, p. 193) Disease Associated HLA Allele Relative Risk** Ankylosing Spondylitis*B27 90 Hereditary HemochromatosisA3/B14 90 Insulin Dependent Diabetes*DR4/DR3 20 Multiple Sclerosis*DR2 5 Myasthenia Gravis*DR3 10 Rheumatoid Arthritis*DR4 10 Systemic Lupus Erythromatosis*DR3 5 NarcolepsyDR2 130 * Autoimmune Disease**Percent of Patients with Allele Divided by Percent of Non-Affected Persons with this Allele HLASick See Table 7-4, Kuby 4th edition, p. 194

63. Limited diversity in HLA gene polymorphism due to breeding bottle-neck in recent past leaves cheetahs exceptionally susceptible to viral infections. (6 th Ed. P. 206)

64. MHC Genes and Tissue Typing: Not covered in BIO 447 in 2012

65. Tissue-Typing Among Siblings: Why Are Blood Relatives More Likely to Be Compatible Donors? Mom's Haplotypes (A,B,C, DP, DQ, DR) Dad's Haplotypes (A,B,C,DP, DQ, DR) 1,2,3,4,5,6 7,8,9,10,11,12 13,14,15,16,17,18 19,20,21,22,23,24 Children's Possible Haplotypes: Inherit from Mom: 1,2,3,4,5,61,2,3,4,5,67,8,9,10,11,127,8,9,10,11,12 Inherit from Dad: 13 to 1819 to 24 13 to 1819 to 24 25% of the Off-Spring Likely to be HLA Haplotypes: 1,2,3,4,5,6 + 13,14,15,16,17,18 1,2,3,4,5,6 + 19,20,21,22,23,24 7,8,9,10,11,12 + 13,14,15,16,17,18 7,8,9,10,11,12 + 19,20,21,22,23,24 SibMatch

66. Illustration of HLA Polymorphism, Linkage Disequilibrium,and Tissue-Typing in Humans Class I Loci Class II Loci HLA-A HLA-B HLA-C DP DQ DR 1 11 1 1 1 2 22 2 2 2 3 33 3 3 3 4 44 4 4 4 5 55 5 5 5 Example HLA-Types for Four Individuals 2 41 5 2 4 3 15 5 1 5 2 41 3 1 2 1 41 2 5 1 HLATyping

67. Tissue-Typing Among Siblings: Why Are Blood Relatives More Likely to Be Compatible Donors? (SibMatch for Dummies version) Mom's Parents (Nigel and Millicent) Dad's Parents (Duwayne and Loquida) Nigel's Haplotypes: Nigel & W Duwayne's Haplotypes: Duwayne & Y Millicent Haplotypes: Millicent & X Loquida's Haplotypes: Loquida & Z Mom's Possible Haplotypes:Dad's Possible Haplotypes: Nigel & MillicentDuwayne & Loquida Nigel & XDuwayne & Z W & Millicent Y and Loquida W & XY & Z Your Possible Haplotypes: Assume Your Mom got the Nigel & Millicent Haplotypes Assume Your Dad got the Duwayne & Loquida Haplotypes Inherit from Mom: Nigel or Millicent Inherit from Dad: Duwayne or Loquida 25% of the Off-Spring Likely to be these Haplotypes: Nigel + Duwayne Nigel + Loquida Millicent+ Duwayne Millicent + Loquida

68. Figure 8-11, Kuby 6 th Edition, p.202, (Selected Part) D Class I Locus KIA & IB Major Class II Loci Class I Locus Mouse H2 Complex Detail

69. I am here! (Testing your XR Transmitter) 1.Yes 2.No Menu Send User Data Hit Enter Get: New channel # Hit: Enter Get: Receiver Found WAIT!!! Get Presentation Mode WAIT! Get Blank Screen Now enter the number “1” for your response Get: Your answer; 1A and a Check-mark Your receiver will appear on the screen 0 of 5 Response Grid

70. H2 Haplotypes of some mouse strains

71. Presentation of Exo-Antigen (Class II) See Figure 8-8, Kuby, 4th Edition, p. 209

72. Presentation of Endogenous Antigen (Class I ) See Figure 8-8, Kuby, 4th Edition, p. 209