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Genetics in the Generation of Antibody Diversity

Genetics in the Generation of Antibody Diversity. Folder Title: AbGenes. Updated: September 29, 2011. GeneTitl. Questions About B-Cell Responses Biochemical Questions: How can Immunoglobulins recognize so many different epitopes ? What does an antibody protein look like?

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Genetics in the Generation of Antibody Diversity

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  1. Genetics in the Generation of Antibody Diversity Folder Title: AbGenes Updated: September 29, 2011 GeneTitl

  2. Questions About B-Cell Responses • Biochemical Questions: • How can Immunoglobulins recognize so many different epitopes? • What does an antibody protein look like? • How do specific antibodies work as proteins binding specific antigenic determinants?

  3. Representation of Sequence Comparisons Among Light Chains from Antibodies with Three Different Antigen Specificities H3N-Ser-Val-Ile-Thr-Gly-Gly-Tyr-Ala... Thr-Glu-Ala-Val-Tyr-Ser-Met-COO- H3N-Ser-Ile-Met-Thr-Arg-Leu-Tyr-Gly..Thr-Glu-Ala-Val-Tyr-Ser-Met-COO- H3N-Thr-Gly-Gly-Thr-Lys-Leu-Tyr-Ile..Thr-Glu-Ala-Val-Tyr-Ser-Met-COO- Variable Amino Terminal HalfConserved Carboxyl Terminal Half (Positions 1 to 107) (Positions 108 - 214) LiteComp

  4. See Figure 4-6, Kuby 6th Edition p. 85

  5. XR: Menu Send User Data Hit Enter; Get: New channel # Hit: Enter; Get: Receiver Found WAIT!!! Get Presentation Mode WAIT! Get Blank Screen Testing doing a “Fill-in-the Blank” Question:Please send in your last name up to the first five letters: For XR: Press any key to activate your transmitter. A string of letters should appear Select letter using Left or Right Arrows (keys 1A or 3C) SEL key will select the chosen letter Use CLR key to erase a letter you don’t want After response is complete press enter Your screen will read “Your Answer”: (Your Entry) with a check mark ____ _____ _____ _____ _____ Non-Response Grid 0 of 103

  6. Questions About B-Cell Responses • Biochemical Questions: • How can Immunoglobulins recognize so many different epitopes? • What does an antibody protein look like? • How do specific antibodies work as proteins binding specific antigenic determinants? • Genetic Question: • How can this diversity of structure leading to enormous diversity of function be coded and controlled by a very limited host genome?

  7. Problems for Genetics in Generating Antibody Sequence Diversity Vast Sequence Diversity- Encoded by Very Limit Genome Heavy and Light Chain Sequence Variations- Almost Exclusively in only one region Exactly the same V-region sequences- End up on different C-region Isotypes GenQuest

  8. Myeloma Cell DNA

  9. Myeloma Cell Conclusions

  10. Embryonic Mouse Cell DNA

  11. Immunology and Phone Numbers 315-443-1870 212-345-1775 478-367-8903 537-503-2078 409-159-6309 610-970-3970 934-620-8122 909-603-7023 800-620-6021 704-590-5307 703-725-0153 207-502-6671 435-431-0890 412-830-0048 740-592-1954 307-620-4450 490-501-5672 601-909-7002 554-891-7712 335-592-0944 Phones

  12. 315- 212- 478- 537- 409- 610- 934- 909- 800- 704- 703- 207- 435- 412- 740- 307- 490- 601- 554- 335- Immunology and Phone Numbers -443-1870 -345-1775 -367-8903 -503-2078 -159-6309 -970-3970 -620-8122 -603-7023 -620-6021 -590-5307 -725-0153 -502-6671 -431-0890 -830-0048 -592-1954 -620-4450 -501-5672 -909-7002 -891-7712 -592-0944 SplicePhones

  13. 315- 212- 478- 537- 409- 610- 934- 909- 800- 704- 703- 207- 435- 412- 740- 307- 490- 601- 554- 335- -443- -345- -367- -503- -159- -970- -620- -603- -620- -590- -725- -502- -431- -830- -592- -620- -501- -909- -891- -592- Immunology and Phone Numbers -1870 -1775 -8903 -2078 -6309 -3970 -8122 -7023 -6021 -5307 -0153 -6671 -0890 -0048 -1954 -4450 -5672 -7002 -7712 -0944 SplicePhones2

  14. Kappa Chain DNA: Vk’s

  15. Kappa Chain: K-J- C

  16. Complete Kappa Chain

  17. Heavy Chain Dna: VH’s, C-Genes

  18. Heavy Chain DNA : All C-Genes

  19. Complete Heavy Chain

  20. Please put away all notes and devices other than your XR transmitter. No consulting is permitted. This is a graded response question.

  21. In the previous slide there were three different regions of genetic information shown that are combined to make antibody chains. “V” stands for variable.What do “J” and “C” stand for?(You can shorten the 2 responses if you run out of characters) Response Grid 0 of 103

  22. When exactly the same V-region specificity of an antibody is genetically linked to different C-region genes (mu, delta, gamma, alpha, or epsilon) that is called class or isotype___ ___ ___ ___ ___or___ ___ ___ ___ ___ ___ ____.switching. Response Grid 0 of 103

  23. From Kuby Immunology, 4th Ed. Upper Half of Table: Human Data Lower Half (not shown): Mouse Data Additional sequence diversity estimated at several orders of magnitude (i.e. perhaps 1000-fold) is generated by junctional flexibility, P-region nucleotide addition, N-region nucleotide addition, and somatic hypermutation. This allows for an estimated 100 million to a billion different possible antibody seuquences in humans. AbDivers

  24. On a scale of 5 to 1 rate todays AbGenes Presentation5. Too slow. Get moving4. No problem.3. About right. I can figure out what I missed.2. Too fast1. I’m lost!(This is anonymous. We won’t know who you are.)

  25. Save Session with Participant List Responses. Close Session. Reset to Anonymous Polling Open New Session for Anonymous Slide on Scope and Pace of the course so far.

  26. This question will be set to anonymous so no one will know what any specific student says. For the first 10 classes the pace of the course is: • Too slow. We need to move faster and cover more about Immunology. • About right. I can keep up Ok but it is a challenge. • Way too fast. Slow down! We’ll figure out later what you don’t get to cover in class. Response Grid 0 of 105

  27. I certify that I am here • True Response Grid 0 of 105

  28. B-Cell Differenatiation

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