Introduction to Immunity Antibody Function & Diversity

1. GMS 6006 - Fundamentals of Immunology & Microbiology Spring 2006 February 15, 2006 Introduction to Immunity Antibody Function & Diversity Wayne T. McCormack, Ph.D. R1-102, 392-7413, mccormac@pathology.ufl.edu Introduction: The goal of these two lectures is to provide an overview of the immune system and how it operates.

2. Antibody Gene Rearrangement Why in the world are we considering antibodies now? I thought this section was about molecular biology . . . For further reading: “Immunobiology - The Immune System in Health & Disease”, Janeway et al ., 6th edition, 2005. For the basics: GMS 6006 – Fundamentals of Immunology & Microbiology For the whole story: GMS 6140 “Principles of Immunology” = GMS 6031/6032/6033 immunology advanced modules September 30, 2005 Wayne T. McCormack, Ph.D. PLEASE REVIEW!

3. “ Immunobiology - The Immune System in Health & Disease”, Janeway et al ., 6th edition, 2005 All textbook figures herein identified by Fig. number © Garland Science

5. Monday, October 29, 2001 West Nile virus spreads throughout Florida, here to stay By MIRANDA LEITSINGER Associated Press Writer MIAMI (AP) - Chevy could hardly stand on his own. He didn't eat, he wasn't responding to his owners and he was dragging his hind quarters. The 9-year-old horse, a Palomino Appaloosa that has won medals for sliding stops and spins in equestrian competitions, could barely walk. ''It was so hard to watch ... to see one reduced to this,'' said owner Sheila Anderson, of Coral Springs. ''I just cried and cried.'' Chevy, a golden-colored horse with a white mane and tail, was the first confirmed case in Broward County of the West Nile virus. As of last week, 51 of Florida's 67 counties from the Panhandle to the Keys were under a medical alert for the Health experts expected the number to keep increasing …...

6. Thursday, October 18, 2001 More than 30 on Capitol Hill test positive for anthrax By TODD S. PURDUM and ALISON MITCHELL The New York Times WASHINGTON - Concern about biological terrorism heightened Wednesday as preliminary tests showed that more than 30 workers on Capitol Hill were exposed to anthrax spores from the contaminated letter sent to the office of the Senate majority leader. Officials also disclosed that early testing has shown that the anthrax samples found in media offices in New York and Florida were of the same strain. While there was no evidence that anyone has yet been infected with what Attorney General John Ashcroft called a ''virulent, strong, very serious'' form of powdery anthrax that was found in a letter Monday in the office of Sen. Tom Daschle, D-S.D., word that more than half the workers in his suite were exposed prompted the closure of all six House and Senate office buildings for further screening …...

7. What disease does this man have?

8. December 29. 2002 UF scientists: Hold off on smallpox vaccine By DIANE CHUN Sun Staff Writer The UF researchers say mass vaccinations present more risks than benefits. Beginning in January, county and state public health officials will begin to carry out the Bush administration's plan to vaccinate 10.5 million medical personnel and emergency responders against smallpox. Should the federal government decide to proceed with the final stage of those plans, scheduled for sometime in 2004, as many as 10 million Floridians might be eligible for voluntary vaccination against smallpox. The question will then become: Should they get vaccinated? ……

9. February 28. 2003 3 have reaction to smallpox shot By DIANE CHUN Sun Staff Writer Officials with the state Department of Health reported that three additional health care workers have apparently had adverse reactions to the smallpox vaccine they received as part of Operation Vaccination. Thursday's announcement came slightly more than a week after the focus of the state's effort shifted from health department workers to groups drawn from hospitals throughout Florida. A total of 1,186 people had received the vaccine as of Monday. One earlier reaction had been reported. ……

11. Are there any other medical conditions in which something “foreign” resides in your body? Is an immune response always beneficial? c.2000 G.Caviness, BioGrafix

13. Edward Jenner’s experiment (1796) material from cowpox lesion of milkmaid village boy susceptible to smallpox inoculate with cowpox wait 2 months inoculate with smallpox child recovers from cowpox child does not develop smallpox

17. Spaces of the Body That Need to Be Monitored and Defended Major classes of pathogens: bacteria viruses protozoa, fungi, parasites Fig. 10-4

19. Concept Map of “Immunity” Immunity Innate (phagocytes) Adaptive (lymphocytes) Surface (skin, mucous membrane) Subsurface Humoral (Ab) Cellular (CMI) Systemic (IgM, IgG) Mucosal (IgA) Cells (PMN; Macrophages; NK) Proteins (Complement [C ′ ])

25. Many bacteria activate phagocytosis and trigger inflammation

32. B cell-rich T cell-rich Lymph node Fig. 1-8

33. Spleen Peyer’s patches Fig. 1-9,1-10

35. Lymphocytes are activated by antigen & Ag-specific cells proliferate in a clonal fashion Fig. 1-13 B cells respond to native Ag T cells respond to “processed & presented” Ag

38. Concept Map of “Immunity” Immunity Innate (phagocytes) Adaptive (lymphocytes) Surface (skin, mucous membrane) Subsurface Humoral (Ab) Cellular (CMI) Systemic (IgM, IgG) Mucosal (IgA) Cells (PMN; Macrophages; NK) Proteins (Complement [C’])

39. Concept Map of “Immunity” Immunity second half Innate (phagocytes) Adaptive (lymphocytes) Surface (skin, mucous membrane) Subsurface Humoral (Ab) Cellular (CMI) Systemic (IgM, IgG) Mucosal (IgA) Cells (PMN; Macrophages; NK) Proteins (Complement [C’])

41. von Behring and Kitasato experiment (1892) isolate serum from blood healthy rabbit diphtheria toxoid vaccinated rabbit diphtheria-resistant rabbit rabbit lives rabbit dies x healthy rabbit healthy rabbit virulent diphtheria

42. Metchnikoff’s “anti-korperen” Microorganisms are engulfed & digested by phagocytic cells (macrophages) Substances in the blood, called or anti-korperen ( antibodies ) enhanced phagocytosis

44. Antibody Structure: The Puzzle solved (mostly) by Porter, Edelman, Nisonoff in 1959-1964 Experimental approach : Break Ab molecules into smaller fragments Reducing agent (break S-S bonds) 2 heavy chains (55 kD) + 2 light chains (25 kD) Fab: fragment antigen-binding Fc: fragment crystalizable

45. ANTIBODY STRUCTURE An antibody molecule is composed of two identical Ig heavy chains (H) and two identical light chains (L), each with a variable region (V) & constant region (C). Amino acid sequences were determined from myeloma proteins. Fig. 1-17,1-16

46. Bence-Jones proteins (myeloma proteins) = Ig light chains found in the urine of patients with multiple myeloma Serum protein electrophoresis multiple myeloma normal serum Clinical terms: hypergammaglobulinemia hypogammaglobulinemia agammaglobulinemia monoclonal gammopathy

48. Monoclonal Antibodies & Hybridomas Kohler & Milstein (1974) Fig. A-14

49. Fig. 3-5,3-7

50. Fig. 3-8,3-9

51. Fig. 3-10

52. The problem of antigen receptor diversity Each lymphocyte has a unique receptor Ig for B cells, TCR for T cells How are antigen receptors with a virtually infinite range of specificities encoded by a finite number of genes? Germline vs . somatic diversity Tonegawa discovered in 1976 that antibody genes are encoded as gene segments , and must undergo gene rearrangement during lymphocyte development. During the next ~10 years it was discovered that the V regions of all germline Ig and TCR genes are encoded by families of gene segments. Fig. 1-18

54. Classes Subclasses Ig isotypes Fig. 4-18

58. Fig. 2-18

61. DISTRIBUTION OF Ig ISOTYPES Fig. 9-22

62. Fig. 4-17

63. FUNCTIONS OF Ig ISOTYPES Fig. 9-1

64. Fig. 9-24,9-25

65. Fig. 9-31,9-32

66. Course of Typical Acute Infection Fig. 10-1

67. Kinetics of Antibody Response Fig. 1-20

68. The affinity as well as the amount of antibody increases with repeated immunization Questions? Fig. 10-31

69. Concept Map of “Immunity” Immunity Lymphocyte development & tolerance T cell recognition of Ag Friday Innate (phagocytes) Adaptive (lymphocytes) Surface (skin, mucous membrane) Subsurface Humoral (Ab) Cellular (CMI) Systemic (IgM, IgG) Mucosal (IgA) Cells (PMN; Macrophages; NK) Proteins (Complement [C’])

70. Concept Map of “Immunity” Immunity Humoral immune response T cell-mediated immunity Monday Eric Sobel, M.D., Ph.D. Innate (phagocytes) Adaptive (lymphocytes) Surface (skin, mucous membrane) Subsurface Humoral (Ab) Cellular (CMI) Systemic (IgM, IgG) Mucosal (IgA) Cells (PMN; Macrophages; NK) Proteins (Complement [C’])